Prmt5 inhibitors and uses thereof

ABSTRACT

Described herein are compounds of Formula (I), pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof. Compounds of the present invention are useful for inhibiting PRMT5 activity. Methods of using the compounds for treating PRMT5-mediated disorders are also described.

RELATED APPLICATIONS

This application claims priority under 35 U.S.C. §119(e) to U.S.provisional patent application, U.S. Ser. No. 61/785,095, filed Mar. 14,2013, and to U.S. provisional patent application, U.S. Ser. No.61/745,494, filed Dec. 21, 2012, the entire contents of each of whichare incorporated herein by reference.

BACKGROUND OF THE INVENTION

Epigenetic regulation of gene expression is an important biologicaldeterminant of protein production and cellular differentiation and playsa significant pathogenic role in a number of human diseases.

Epigenetic regulation involves heritable modification of geneticmaterial without changing its nucleotide sequence. Typically, epigeneticregulation is mediated by selective and reversible modification (e.g.,methylation) of DNA and proteins (e.g., histones) that control theconformational transition between transcriptionally active and inactivestates of chromatin. These covalent modifications can be controlled byenzymes such as methyltransferases (e.g., PRMT5), many of which areassociated with specific genetic alterations that can cause humandisease.

Disease-associated chromatin-modifying enzymes (e.g., PRMT5) play a rolein diseases such as proliferative disorders, metabolic disorders, andblood disorders. Thus, there is a need for the development of smallmolecules that are capable of inhibiting the activity of PRMT5.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

Protein arginine methyltransferase 5 (PRMT5) catalyzes the addition oftwo methyl groups to the two w-guanidino nitrogen atoms of arginine,resulting in w-NG, N′G symmetric dimethylation of arginine (sDMA) of thetarget protein. PRMT5 functions in the nucleus as well as in thecytoplasm, and its substrates include histones, spliceosomal proteins,transcription factors (See e.g., Sun et al., 2011, PNAS 108:20538-20543). PRMT5 generally functions as part of a molecule weightprotein complex. While the protein complexes of PRMT5 can have a varietyof components, they generally include the protein MEP50 (methylosomeprotein 50). In addition, PRMT5 acts in conjunction with cofactor SAM(S-adenosyl methionine).

PRMT5 is an attractive target for modulation given its role in theregulation of diverse biological processes. It has now been found thatcompounds described herein, and pharmaceutically acceptable salts andcompositions thereof, are effective as inhibitors of PRMT5. Suchcompounds have the general Formula (I):

or a pharmaceutically acceptable salt thereof, wherein Ring Z, L_(z),R¹, R²¹, R²², R²³, R²⁴, R^(x), and n are as defined herein.

In some embodiments, pharmaceutical compositions are provided whichcomprise a compound described herein (e.g., a compound of Formula (I)),or a pharmaceutically acceptable salt thereof, and optionally apharmaceutically acceptable excipient.

In certain embodiments, compounds described herein inhibit activity ofPRMT5. In certain embodiments, methods of inhibiting PRMT5 are providedwhich comprise contacting PRMT5 with an effective amount of a compoundof Formula (I), or a pharmaceutically acceptable salt thereof. The PRMT5may be purified or crude, and may be present in a cell, tissue, or asubject. Thus, such methods encompass inhibition of PRMT5 activity bothin vitro and in vivo. In certain embodiments, the PRMT5 is wild-typePRMT5. In certain embodiments, the PRMT5 is overexpressed. In certainembodiments, the PRMT5 is a mutant. In certain embodiments, the PRMT5 isin a cell. In certain embodiments, the PRMT5 is in an animal, e.g., ahuman. In some embodiments, the PRMT5 is in a subject that issusceptible to normal levels of PRMT5 activity due to one or moremutations associated with a PRMT5 substrate. In some embodiments, thePRMT5 is in a subject known or identified as having abnormal PRMT5activity (e.g., overexpression). In some embodiments, a providedcompound is selective for PRMT5 over other methyltransferases. Incertain embodiments, a provided compound is at least about 10-foldselective, at least about 20-fold selective, at least about 30-foldselective, at least about 40-fold selective, at least about 50-foldselective, at least about 60-fold selective, at least about 70-foldselective, at least about 80-fold selective, at least about 90-foldselective, or at least about 100-fold selective relative to one or moreother methyltransferases.

In certain embodiments, methods of altering gene expression in a cellare provided which comprise contacting a cell with an effective amountof a compound of Formula (I), or a pharmaceutically acceptable saltthereof, or a pharmaceutical composition thereof. In certainembodiments, the cell in culture in vitro. In certain embodiments, cellis in an animal, e.g., a human.

In certain embodiments, methods of altering transcription in a cell areprovided which comprise contacting a cell with an effective amount of acompound of Formula (I), or a pharmaceutically acceptable salt thereof,or a pharmaceutical composition thereof. In certain embodiments, thecell in culture in vitro. In certain embodiments, the cell is in ananimal, e.g., a human.

In some embodiments, methods of treating a PRMT5-mediated disorder areprovided which comprise administering to a subject suffering from aPRMT5-mediated disorder an effective amount of a compound describedherein (e.g., a compound of Formula (I)), or a pharmaceuticallyacceptable salt thereof, or a pharmaceutical composition thereof. Incertain embodiments, the PRMT5-mediated disorder is a proliferativedisorder, a metabolic disorder, or a blood disorder. In certainembodiments, compounds described herein are useful for treating cancer.In certain embodiments, compounds described herein are useful fortreating hematopoietic cancer, lung cancer, prostate cancer, melanoma,or pancreatic cancer. In certain embodiments, compounds described hereinare useful for treating a hemoglobinopathy. In certain embodiments,compounds described herein are useful for treating sickle cell anemia.In certain embodiments, compounds described herein are useful fortreating diabetes or obesity.

Compounds described herein are also useful for the study of PRMT5 inbiological and pathological phenomena, the study of intracellular signaltransduction pathways mediated by PRMT5, and the comparative evaluationof new PRMT5 inhibitors.

This application refers to various issued patent, published patentapplications, journal articles, and other publications, all of which areincorporated herein by reference.

Definitions of specific functional groups and chemical terms aredescribed in more detail below. The chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75^(n) Ed., a inside cover, andspecific functional groups are generally defined as described therein.Additionally, general principles of organic chemistry, as well asspecific functional moieties and reactivity, are described in ThomasSorrell, Organic Chemistry, University Science Books, Sausalito, 1999;Smith and March, March's Advanced Organic Chemistry, 5^(th) Edition,John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive OrganicTransformations, VCH Publishers, Inc., New York, 1989; and Carruthers,Some Modern Methods of Organic Synthesis, 3^(rd) Edition, CambridgeUniversity Press, Cambridge, 1987.

Compounds described herein can comprise one or more asymmetric centers,and thus can exist in various isomeric forms, e.g., enantiomers and/ordiastereomers. For example, the compounds described herein can be in theform of an individual enantiomer, diastereomer or geometric isomer, orcan be in the form of a mixture of stereoisomers, including racemicmixtures and mixtures enriched in one or more stereoisomer. Isomers canbe isolated from mixtures by methods known to those skilled in the art,including chiral high pressure liquid chromatography (HPLC) and theformation and crystallization of chiral salts; or preferred isomers canbe prepared by asymmetric syntheses. See, for example, Jacques et al.,Enantiomers, Racemates and Resolutions (Wiley Interscience, New York,1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistryof Carbon Compounds (McGrawHill, NY, 1962); and Wilen, Tables ofResolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ.of Notre Dame Press, Notre Dame, Ind. 1972). The present disclosureadditionally encompasses compounds described herein as individualisomers substantially free of other isomers, and alternatively, asmixtures of various isomers.

It is to be understood that the compounds of the present invention maybe depicted as different tautomers. It should also be understood thatwhen compounds have tautomeric forms, all tautomeric forms are intendedto be included in the scope of the present invention, and the naming ofany compound described herein does not exclude any tautomer form.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds that differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of hydrogen by deuterium ortritium, replacement of ¹⁹F with ¹⁸F, or the replacement of a carbon bya ¹³C- or ¹⁴C-enriched carbon are within the scope of the disclosure.Such compounds are useful, for example, as analytical tools or probes inbiological assays.

The term “aliphatic,” as used herein, includes both saturated andunsaturated, nonaromatic, straight chain (i.e., unbranched), branched,acyclic, and cyclic (i.e., carbocyclic) hydrocarbons. In someembodiments, an aliphatic group is optionally substituted with one ormore functional groups. As will be appreciated by one of ordinary skillin the art, “aliphatic” is intended herein to include alkyl, alkenyl,alkynyl, cycloalkyl, and cycloalkenyl moieties.

When a range of values is listed, it is intended to encompass each valueand sub-range within the range. For example “C₁₋₆ alkyl” is intended toencompass, C₁, C₂, C₃, C₄, C₅, C₆, C₁₋₆, C₁₋₅, C₁₋₄, C₁₋₃, C₁₋₂, C₂₋₆,C₂₋₅, C₂₋₄, C₂₋₃, C₃₋₆, C₃₋₅, C₃₋₄, C₄₋₆, C₄₋₅, and C₅₋₆ alkyl.

“Alkyl” refers to a radical of a straight-chain or branched saturatedhydrocarbon group having from 1 to 20 carbon atoms (“C₁₋₂₀ alkyl”). Insome embodiments, an alkyl group has 1 to 10 carbon atoms (“C₁₋₁₀alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms(“C₁₋₉ alkyl”). In some embodiments, an alkyl group has 1 to 8 carbonatoms (“C₁₋₈ alkyl”). In some embodiments, an alkyl group has 1 to 7carbon atoms (“C₁₋₇ alkyl”). In some embodiments, an alkyl group has 1to 6 carbon atoms (“C₁₋₆ alkyl”). In some embodiments, an alkyl grouphas 1 to 5 carbon atoms (“C₁₋₅ alkyl”). In some embodiments, an alkylgroup has 1 to 4 carbon atoms (“C₁₋₄ alkyl”). In some embodiments, analkyl group has 1 to 3 carbon atoms (“C₁₋₃ alkyl”). In some embodiments,an alkyl group has 1 to 2 carbon atoms (“C₁₋₂ alkyl”). In someembodiments, an alkyl group has 1 carbon atom (“C₁₋₆ alkyl”). In someembodiments, an alkyl group has 2 to 6 carbon atoms (“C₂₋₆ alkyl”).Examples of C₁₋₆ alkyl groups include methyl (C₁), ethyl (C₂), n-propyl(C₃), isopropyl (C₃), n-butyl (C₄), tert-butyl (C₄), sec-butyl (C₄),iso-butyl (C₄), n-pentyl (C₅), 3-pentanyl (C₅), amyl (C₅), neopentyl(C₅), 3-methyl-2-butanyl (C₅), tertiary amyl (C₅), and n-hexyl (C₆).Additional examples of alkyl groups include n-heptyl (C₇), n-octyl (C₈)and the like. In certain embodiments, each instance of an alkyl group isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted alkyl”) or substituted (a “substituted alkyl”) with oneor more substituents. In certain embodiments, the alkyl group isunsubstituted C₁₋₁₀ alkyl (e.g., —CH₃). In certain embodiments, thealkyl group is substituted C₁₋₁₀ alkyl.

In some embodiments, an alkyl group is substituted with one or morehalogens. “Perhaloalkyl” is a substituted alkyl group as defined hereinwherein all of the hydrogen atoms are independently replaced by ahalogen, e.g., fluoro, bromo, chloro, or iodo. In some embodiments, thealkyl moiety has 1 to 8 carbon atoms (“C₁₋₈ perhaloalkyl”). In someembodiments, the alkyl moiety has 1 to 6 carbon atoms (“C₁₋₆perhaloalkyl”). In some embodiments, the alkyl moiety has 1 to 4 carbonatoms (“C₁₋₆ perhaloalkyl”). In some embodiments, the alkyl moiety has 1to 3 carbon atoms (“C₁₋₃ perhaloalkyl”). In some embodiments, the alkylmoiety has 1 to 2 carbon atoms (“C₁₋₂ perhaloalkyl”). In someembodiments, all of the hydrogen atoms are replaced with fluoro. In someembodiments, all of the hydrogen atoms are replaced with chloro.Examples of perhaloalkyl groups include CF₃, CF₂CF₃, CF₂CF₂CF₃, CCl₃,CFCl₂, CF₂Cl, and the like.

“Alkenyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, one or morecarbon-carbon double bonds, and no triple bonds (“C₂₋₂₀ alkenyl”). Insome embodiments, an alkenyl group has 2 to 10 carbon atoms (“C₂₋₁₀alkenyl”). In some embodiments, an alkenyl group has 2 to 9 carbon atoms(“C₂₋₉ alkenyl”). In some embodiments, an alkenyl group has 2 to 8carbon atoms (“C₂₋₈ alkenyl”). In some embodiments, an alkenyl group has2 to 7 carbon atoms (“C₂₋₇ alkenyl”). In some embodiments, an alkenylgroup has 2 to 6 carbon atoms (“C₂₋₆ alkenyl”). In some embodiments, analkenyl group has 2 to 5 carbon atoms (“C₂₋₅ alkenyl”). In someembodiments, an alkenyl group has 2 to 4 carbon atoms (“C₂₋₄ alkenyl”).In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C₂₋₃alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C₂alkenyl”). The one or more carbon-carbon double bonds can be internal(such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples ofC₂₋₄ alkenyl groups include ethenyl (C₂), 1 (C₃), 2-propenyl (C₃),1-butenyl (C₄), 2-butenyl (C₄), butadienyl (C₄), and the like. Examplesof C₂₋₆ alkenyl groups include the aforementioned C₂₋₄ alkenyl groups aswell as pentenyl (C₅), pentadienyl (C₅), hexenyl (C₆), and the like.Additional examples of alkenyl include heptenyl (C₇), octenyl (C₈),octatrienyl (C₈), and the like. In certain embodiments, each instance ofan alkenyl group is independently optionally substituted, e.g.,unsubstituted (an “unsubstituted alkenyl”) or substituted (a“substituted alkenyl”) with one or more substituents. In certainembodiments, the alkenyl group is unsubstituted C₂₋₁₀ alkenyl. Incertain embodiments, the alkenyl group is substituted C₂₋₁₀ alkenyl.

“Alkynyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, one or morecarbon-carbon triple bonds, and optionally one or more double bonds(“C₂₋₂₀ alkynyl”). In some embodiments, an alkynyl group has 2 to 10carbon atoms (“C₂₋₁₀ alkynyl”). In some embodiments, an alkynyl grouphas 2 to 9 carbon atoms (“C₂₋₉ alkynyl”). In some embodiments, analkynyl group has 2 to 8 carbon atoms (“C₂₋₈ alkynyl”). In someembodiments, an alkynyl group has 2 to 7 carbon atoms (“C₂₋₇ alkynyl”).In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C₂₋₆alkynyl”). In some embodiments, an alkynyl group has 2 to 5 carbon atoms(“C₂₋₅ alkynyl”). In some embodiments, an alkynyl group has 2 to 4carbon atoms (“C₂₋₄ alkynyl”). In some embodiments, an alkynyl group has2 to 3 carbon atoms (“C₂₋₃ alkynyl”). In some embodiments, an alkynylgroup has 2 carbon atoms (“C₂ alkynyl”). The one or more carbon-carbontriple bonds can be internal (such as in 2-butynyl) or terminal (such asin 1-butynyl). Examples of C₂₋₄ alkynyl groups include, withoutlimitation, ethynyl (C₂), 1-propynyl (C₃), 2-propynyl (C₃), 1-butynyl(C₄), 2-butynyl (C₄), and the like. Examples of C₂₋₆ alkenyl groupsinclude the aforementioned C₂₋₄ alkynyl groups as well as pentynyl (C₅),hexynyl (C₆), and the like. Additional examples of alkynyl includeheptynyl (C₇), octynyl (C₈), and the like. In certain embodiments, eachinstance of an alkynyl group is independently optionally substituted,e.g., unsubstituted (an “unsubstituted alkynyl”) or substituted (a“substituted alkynyl”) with one or more substituents. In certainembodiments, the alkynyl group is unsubstituted C₂₋₁₀ alkynyl. Incertain embodiments, the alkynyl group is substituted C₂₋₁₀ alkynyl.

“Carbocyclyl” or “carbocyclic” refers to a radical of a nonaromaticcyclic hydrocarbon group having from 3 to 10 ring carbon atoms (“C₃₋₁₀carbocyclyl”) and zero heteroatoms in the nonaromatic ring system. Insome embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms(“C₃₋₈ carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to6 ring carbon atoms (“C₃₋₆ carbocyclyl”). In some embodiments, acarbocyclyl group has 3 to 6 ring carbon atoms (“C₃₋₆ carbocyclyl”). Insome embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms(“C₅₋₁₀ carbocyclyl”). Exemplary C₃₋₆ carbocyclyl groups include,without limitation, cyclopropyl (C₃), cyclopropenyl (C₃), cyclobutyl(C₄), cyclobutenyl (C₄), cyclopentyl (C₅), cyclopentenyl (C₅),cyclohexyl (C₆), cyclohexenyl (C₆), cyclohexadienyl (C₆), and the like.Exemplary C₃₋₈ carbocyclyl groups include, without limitation, theaforementioned C₃₋₆ carbocyclyl groups as well as cycloheptyl (C₇),cycloheptenyl (C₇), cycloheptadienyl (C₇), cycloheptatrienyl (C₇),cyclooctyl (C₈), cyclooctenyl (C₈), bicyclo[2.2.1]heptanyl (C₇),bicyclo[2.2.2]octanyl (C₈), and the like. Exemplary C₃₋₁₀ carbocyclylgroups include, without limitation, the aforementioned C₃₋₈ carbocyclylgroups as well as cyclononyl (C₉), cyclononenyl (C₉), cyclodecyl (C₁₀),cyclodecenyl (C₁₀), octahydro-1H-indenyl (C₉), decahydronaphthalenyl(C₁₀), spiro[4.5]decanyl (C₁₀), and the like. As the foregoing examplesillustrate, in certain embodiments, the carbocyclyl group is eithermonocyclic (“monocyclic carbocyclyl”) or contain a fused, bridged orspiro ring system such as a bicyclic system (“bicyclic carbocyclyl”) andcan be saturated or can be partially unsaturated. “Carbocyclyl” alsoincludes ring systems wherein the carbocyclyl ring, as defined above, isfused with one or more aryl or heteroaryl groups wherein the point ofattachment is on the carbocyclyl ring, and in such instances, the numberof carbons continue to designate the number of carbons in thecarbocyclic ring system. In certain embodiments, each instance of acarbocyclyl group is independently optionally substituted, e.g.,unsubstituted (an “unsubstituted carbocyclyl”) or substituted (a“substituted carbocyclyl”) with one or more substituents. In certainembodiments, the carbocyclyl group is unsubstituted C₃₋₁₀ carbocyclyl.In certain embodiments, the carbocyclyl group is a substituted C₃₋₁₀carbocyclyl.

In some embodiments, “carbocyclyl” is a monocyclic, saturatedcarbocyclyl group having from 3 to 10 ring carbon atoms (“C₃₋₁₀cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 8 ringcarbon atoms (“C₃₋₈ cycloalkyl”). In some embodiments, a cycloalkylgroup has 3 to 6 ring carbon atoms (“C₃₋₆ cycloalkyl”). In someembodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C₅₋₆cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ringcarbon atoms (“C₅₋₁₀ cycloalkyl”). Examples of C₅₋₆ cycloalkyl groupsinclude cyclopentyl (C₅) and cyclohexyl (C₅). Examples of C₃₋₆cycloalkyl groups include the aforementioned C₅₋₆ cycloalkyl groups aswell as cyclopropyl (C₃) and cyclobutyl (C₄). Examples of C₃₋₈cycloalkyl groups include the aforementioned C₃₋₆ cycloalkyl groups aswell as cycloheptyl (C₇) and cyclooctyl (C₈). In certain embodiments,each instance of a cycloalkyl group is independently unsubstituted (an“unsubstituted cycloalkyl”) or substituted (a “substituted cycloalkyl”)with one or more substituents. In certain embodiments, the cycloalkylgroup is unsubstituted C₃₋₁₀ cycloalkyl. In certain embodiments, thecycloalkyl group is substituted C₃₋₁₀ cycloalkyl.

“Heterocyclyl” or “heterocyclic” refers to a radical of a 3 to10-membered non aromatic ring system having ring carbon atoms and 1 to 4ring heteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“3-10 membered heterocyclyl”). Inheterocyclyl groups that contain one or more nitrogen atoms, the pointof attachment can be a carbon or nitrogen atom, as valency permits. Aheterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”)or a fused, bridged or spiro ring system such as a bicyclic system(“bicyclic heterocyclyl”), and can be saturated or can be partiallyunsaturated. Heterocyclyl bicyclic ring systems can include one or moreheteroatoms in one or both rings. “Heterocyclyl” also includes ringsystems wherein the heterocyclyl ring, as defined above, is fused withone or more carbocyclyl groups wherein the point of attachment is eitheron the carbocyclyl or heterocyclyl ring, or ring systems wherein theheterocyclyl ring, as defined above, is fused with one or more aryl orheteroaryl groups, wherein the point of attachment is on theheterocyclyl ring, and in such instances, the number of ring memberscontinue to designate the number of ring members in the heterocyclylring system. In certain embodiments, each instance of heterocyclyl isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted heterocyclyl”) or substituted (a “substitutedheterocyclyl”) with one or more substituents. In certain embodiments,the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. Incertain embodiments, the heterocyclyl group is substituted 3-10 memberedheterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 membered nonaromaticring system having ring carbon atoms and 1-4 ring heteroatoms, whereineach heteroatom is independently selected from nitrogen, oxygen, andsulfur (“5-10 membered heterocyclyl”). In some embodiments, aheterocyclyl group is a 5-8 membered nonaromatic ring system having ringcarbon atoms and 1-4 ring heteroatoms, wherein each heteroatom isindependently selected from nitrogen, oxygen, and sulfur (“5-8 memberedheterocyclyl”). In some embodiments, a heterocyclyl group is a 5-6membered nonaromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”). In someembodiments, the 5-6 membered heterocyclyl has 1-3 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has 1-2 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has one ring heteroatomselected from nitrogen, oxygen, and sulfur.

Exemplary 3-membered heterocyclyl groups containing one heteroatominclude, without limitation, azirdinyl, oxiranyl, and thiorenyl.Exemplary 4-membered heterocyclyl groups containing one heteroatominclude, without limitation, azetidinyl, oxetanyl, and thietanyl.Exemplary 5-membered heterocyclyl groups containing one heteroatominclude, without limitation, tetrahydrofuranyl, dihydrofuranyl,tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl,and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groupscontaining two heteroatoms include, without limitation, dioxolanyl,oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-memberedheterocyclyl groups containing three heteroatoms include, withoutlimitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary6-membered heterocyclyl groups containing one heteroatom include,without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl,and thianyl. Exemplary 6 heterocyclyl groups containing two heteroatomsinclude, without limitation, piperazinyl, morpholinyl, dithianyl, anddioxanyl. Exemplary 6-membered heterocyclyl groups containing twoheteroatoms include, without limitation, triazinanyl. Exemplary7-membered heterocyclyl groups containing one heteroatom include,without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary8-membered heterocyclyl groups containing one heteroatom include,without limitation, azocanyl, oxecanyl, and thiocanyl. Exemplary5-membered heterocyclyl groups fused to a C₁₋₆ aryl ring (also referredto herein as a 5,6-bicyclic heterocyclic ring) include, withoutlimitation, indolinyl, isoindolinyl, dihydrobenzofuranyl,dihydrobenzothienyl, benzoxazolinonyl, and the like. Exemplary6-membered heterocyclyl groups fused to an aryl ring (also referred toherein as a 6,6-bicyclic heterocyclic ring) include, without limitation,tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.

“Aryl” refers to a radical of a monocyclic or polycyclic (e.g., bicyclicor tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 itelectrons shared in a cyclic array) having 6-14 ring carbon atoms andzero heteroatoms provided in the aromatic ring system (“C₆₋₁₄ aryl”). Insome embodiments, an aryl group has six ring carbon atoms (“C₁₋₆ aryl”;e.g., phenyl). In some embodiments, an aryl group has ten ring carbonatoms (“C₁₀ aryl”; e.g., naphthyl such as 1-naphthyl and 2-naphthyl). Insome embodiments, an aryl group has fourteen ring carbon atoms (“C₁₋₄aryl”; e.g., anthracyl). “Aryl” also includes ring systems wherein thearyl ring, as defined above, is fused with one or more carbocyclyl orheterocyclyl groups wherein the radical or point of attachment is on thearyl ring, and in such instances, the number of carbon atoms continue todesignate the number of carbon atoms in the aryl ring system. In certainembodiments, each instance of an aryl group is independently optionallysubstituted, e.g., unsubstituted (an “unsubstituted aryl”) orsubstituted (a “substituted aryl”) with one or more substituents. Incertain embodiments, the aryl group is unsubstituted C₆₋₁₄ aryl. Incertain embodiments, the aryl group is substituted C₆₋₁₄ aryl.

“Heteroaryl” refers to a radical of a 5-10 membered monocyclic orbicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 it electronsshared in a cyclic array) having ring carbon atoms and 1-4 ringheteroatoms provided in the aromatic ring system, wherein eachheteroatom is independently selected from nitrogen, oxygen and sulfur(“5-10 membered heteroaryl”). In heteroaryl groups that contain one ormore nitrogen atoms, the point of attachment can be a carbon or nitrogenatom, as valency permits. Heteroaryl bicyclic ring systems can includeone or more heteroatoms in one or both rings. “Heteroaryl” includes ringsystems wherein the heteroaryl ring, as defined above, is fused with oneor more carbocyclyl or heterocyclyl groups wherein the point ofattachment is on the heteroaryl ring, and in such instances, the numberof ring members continue to designate the number of ring members in theheteroaryl ring system. “Heteroaryl” also includes ring systems whereinthe heteroaryl ring, as defined above, is fused with one or more arylgroups wherein the point of attachment is either on the aryl orheteroaryl ring, and in such instances, the number of ring membersdesignates the number of ring members in the fused (aryl/heteroaryl)ring system. Bicyclic heteroaryl groups wherein one ring does notcontain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and thelike) the point of attachment can be on either ring, e.g., either thering bearing a heteroatom (e.g., 2-indolyl) or the ring that does notcontain a heteroatom (e.g., 5-indolyl).

In some embodiments, a heteroaryl group is a 5-10 membered aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-8 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-6 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In someembodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heteroaryl has 1-2 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selectedfrom nitrogen, oxygen, and sulfur. In certain embodiments, each instanceof a heteroaryl group is independently optionally substituted, e.g.,unsubstituted (“unsubstituted heteroaryl”) or substituted (“substitutedheteroaryl”) with one or more substituents. In certain embodiments, theheteroaryl group is unsubstituted 5-14 membered heteroaryl. In certainembodiments, the heteroaryl group is substituted 5-14 memberedheteroaryl.

Exemplary 5-membered heteroaryl groups containing one heteroatominclude, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary5-membered heteroaryl groups containing two heteroatoms include, withoutlimitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, andisothiazolyl. Exemplary 5-membered heteroaryl groups containing threeheteroatoms include, without limitation, triazolyl, oxadiazolyl, andthiadiazolyl. Exemplary 5-membered heteroaryl groups containing fourheteroatoms include, without limitation, tetrazolyl. Exemplary6-membered heteroaryl groups containing one heteroatom include, withoutlimitation, pyridinyl. Exemplary 6-membered heteroaryl groups containingtwo heteroatoms include, without limitation, pyridazinyl, pyrimidinyl,and pyrazinyl. Exemplary 6-membered heteroaryl groups containing threeor four heteroatoms include, without limitation, triazinyl andtetrazinyl, respectively. Exemplary 7-membered heteroaryl groupscontaining one heteroatom include, without limitation, azepinyl,oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groupsinclude, without limitation, indolyl, isoindolyl, indazolyl,benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl,benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl,benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl,indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groupsinclude, without limitation, naphthyridinyl, pteridinyl, quinolinyl,isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

“Partially unsaturated” refers to a group that includes at least onedouble or triple bond. The term “partially unsaturated” is intended toencompass rings having multiple sites of unsaturation, but is notintended to include aromatic groups (e.g., aryl or heteroaryl groups) asherein defined. Likewise, “saturated” refers to a group that does notcontain a double or triple bond, i.e., contains all single bonds.

In some embodiments, aliphatic, alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aryl, and heteroaryl groups, as defined herein, areoptionally substituted (e.g., “substituted” or “unsubstituted”aliphatic, “substituted” or “unsubstituted” alkyl, “substituted” or“unsubstituted” alkenyl, “substituted” or “unsubstituted” alkynyl,“substituted” or “unsubstituted” carbocyclyl, “substituted” or“unsubstituted” heterocyclyl, “substituted” or “unsubstituted” aryl or“substituted” or “unsubstituted” heteroaryl group). In general, the term“substituted”, whether preceded by the term “optionally” or not, meansthat at least one hydrogen present on a group (e.g., a carbon ornitrogen atom) is replaced with a permissible substituent, e.g., asubstituent which upon substitution results in a stable compound, e.g.,a compound which does not spontaneously undergo transformation such asby rearrangement, cyclization, elimination, or other reaction. Unlessotherwise indicated, a “substituted” group has a substituent at one ormore substitutable positions of the group, and when more than oneposition in any given structure is substituted, the substituent iseither the same or different at each position. The term “substituted” iscontemplated to include substitution with all permissible substituentsof organic compounds, including any of the substituents described hereinthat results in the formation of a stable compound. The presentdisclosure contemplates any and all such combinations in order to arriveat a stable compound. For purposes of this disclosure, heteroatoms suchas nitrogen may have hydrogen substituents and/or any suitablesubstituent as described herein which satisfy the valencies of theheteroatoms and results in the formation of a stable moiety.

Exemplary carbon atom substituents include, but are not limited to,halogen, —CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(aa), —ON(R^(bb))₂,—N(R^(bb))₂, —N(R^(bb))₃ ⁺X⁻, —N(OR^(cc))R^(bb), —SH, —SR^(aa),—SSR^(cc), —C(═O)R^(aa), —CO₂H, —CHO, —C(OR^(cc))₂, —CO₂R^(aa),—OC(═O)R^(aa), —OCO₂R^(aa), —C(═O)N(R^(bb))₂, —OC(═O)N(R^(bb))₂,—NR^(bb)C(═O)R^(aa), —NR^(bb)CO₂R^(aa), —NR^(bb)C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —OC(═NR^(bb))R^(aa),—OC(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂, —OC(═NR^(bb))N(R^(bb))₂,—NR^(bb)C(═NR^(bb))N(R^(bb))₂, —C(═O)NR^(bb)SO₂R^(aa),—NR^(bb)SO₂R^(aa), —SO₂N(R^(bb))₂, —SO₂R^(aa), —SO₂OR, —OSO₂R^(aa),—S(═O)R^(aa), —OS(═O)R^(aa), —Si(R^(aa))₃, —OSi(R^(aa))₃,—C(═S)N(R^(bb))₂, —C(═O)SR^(aa), —C(═S)SR^(aa), —SC(═S)SR^(aa),—SC(═O)SR^(aa), —OC(═O)SR^(aa), —SC(═O)OR^(aa), —SC(═O)R^(aa),—P(═O)₂R^(aa), —OP(═O)₂R^(aa), —P(═O)(R^(aa))₂, —OP(═O)(R^(aa))₂,—OP(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, —OP(═O)₂N(R^(bb))₂,—P(═O)(NR^(bb))₂, —OP(═O)(NR^(bb))₂, —NR^(bb)P(═O)(OR^(cc))₂,—NR^(bb)P(═O)(NR^(bb))₂, —P(R)₂, —P(R^(cc))₃, —OP(R^(cc))₂,—OP(R^(cc))₃, —B(R^(aa))₂, —B(OR^(cc))₂, —BR^(aa)(OR^(cc)), C₁₋₁₀ alkyl,C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ carbocyclyl,3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5R^(dd) groups;

or two geminal hydrogens on a carbon atom are replaced with the group═O, ═S, ═NN(R^(bb))₂, ═NNR^(bb)C(═O)R^(aa), ═NNR^(bb)C(═O)OR^(aa),═NNR^(bb)S(═O)₂R^(aa), ═NR^(bb), or ═NOR^(cc);

each instance of R^(aa) is, independently, selected from C₁₋₁₀ alkyl,C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ carbocyclyl,3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl, ortwo R^(aa) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(bb) is, independently, selected from hydrogen, —OH,—OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂,—SO₂R^(cc), —SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc),—C(═S)SR^(cc), —P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂,—P(═O)(NR^(cc))₂, C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, or two R^(bb) groups are joined to form a 3-14membered heterocyclyl or 5-14 membered heteroaryl ring, wherein eachalkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroarylis independently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(cc) is, independently, selected from hydrogen, C₁₀alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups are joined to form a 3-14 memberedheterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl,alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl isindependently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(dd) is, independently, selected from halogen, —CN,—NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(ee), —ON(R^(ff))₂, —N(R^(ff))₂,—N(R^(ff))₃ ⁺X⁻, —N(OR^(ee))R^(ff), —SH, —SR^(ee), —SSR^(ee),—C(═O)R^(ee), —CO₂H, —CO₂R^(ee), —OC(═O)R^(ee), —OCO₂R^(ee),—C(═O)N(R^(ff))₂, —OC(═O)N(R^(ff))₂, —NR^(ff)C(═O)R^(ee),—NR^(ff)C₂R^(ee), —NR^(ff)C(═O)N(R^(ff))₂, —C(═NR^(ff))OR^(ee),—OC(═NR^(ff))R^(ee), —OC(═NR^(ff))OR^(ee), —C(═NR^(ff))N(R^(ff))₂,—OC(═NR^(ff))N(R^(ff))₂, —NR^(ff)C(═NR^(ff))N(R^(ff))₂,—NR^(ff)SO₂R^(ee), SO₂N(R^(ff))₂, —SO₂R^(ee), —SO₂OR^(ee), —OSO₂R^(ee),—S(═O)R^(ee), —Si(R^(ee))₃, —OSi(R^(ee))₃, —C(═S)N(R^(ff))₂,—C(═O)SR^(ee), —C(═S)SR^(ee), —SC(═S)SR^(ee), —P(═O)₂R^(ee),—P(═O)(R^(ee))₂, —OP(═O)(R^(ee))₂, —OP(═O)(OR^(ee))₂, C₁₋₄ alkyl, C₁₋₆perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, 3-10membered heterocyclyl, C₆₋₁₀ aryl, 5-10 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups, or two geminal R^(dd) substituents can be joined to form ═O or═S;

each instance of R^(ee) is, independently, selected from C₁₋₆ alkyl,C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, C₆₋₁₀aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups; each instance of R^(ff) is, independently, selected fromhydrogen, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₃₋₁₀ carbocyclyl, 3-10 membered heterocyclyl, C₆₋₁₀ aryl and 5-10membered heteroaryl, or two R^(ff) groups are joined to form a 3-14membered heterocyclyl or 5-14 membered heteroaryl ring, wherein eachalkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroarylis independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups; and

each instance of R^(gg) is, independently, halogen, —CN, —NO₂, —N₃,—SO₂H, —SO₃H, —OH, —OC₁₋₆ alkyl, —ON(C₁₋₆ alkyl)₂, —N(C₁₋₆ alkyl)₂,—N(C₁₋₆ alkyl)₃ ⁺X⁻, —NH(C₁₋₆ alkyl)₂ ⁺X⁻, —NH₂(C₁₋₆ alkyl)⁺X⁻, —NH₃⁺X⁻, —N(OC₁₋₆ alkyl)(C₁₋₆ alkyl), —N(OH)(C₁₋₆ alkyl), —NH(OH), —SH,—SC₁₋₆ alkyl, —SS(C₁₋₆ alkyl), —C(═O)(C₁₋₆ alkyl), —CO₂H, —CO₂(C₁₋₆alkyl), —OC(═O)(C₁₋₆ alkyl), —OCO₂(C₁₋₆ alkyl), —C(═O)NH₂, —C(═O)N(C₁₋₆alkyl)₂, —OC(═O)NH(C₁₋₆ alkyl), —NHC(═O)(C₁₋₆ alkyl), —N(C₁₋₆alkyl)C(═O)(C₁₋₆ alkyl), —NHCO₂(C₁₋₆ alkyl), —NHC(═O)N(C₁₋₆ alkyl)₂,—NHC(═O)NH(C₁₋₆ alkyl), —NHC(═O)NH₂, —C(═NH)O(C₁₋₆ alkyl), —OC(═NH)(C₁₋₆alkyl), —OC(═NH)OC₁₋₆ alkyl, —C(═NH)N(C₁₋₆ alkyl)₂, —C(═NH)NH(C₁₋₆alkyl), —C(═NH)NH₂, —OC(═NH)N(C₁₋₆ alkyl)₂, —OC(NH)NH(C₁₋₆ alkyl),—OC(NH)NH₂, —NHC(NH)N(C₁₋₆ alkyl)₂, —NHC(═NH)NH₂, —NHSO₂(C₁₋₆ alkyl),—SO₂N(C₁₋₆ alkyl)₂, —SO₂NH(C₁₋₆ alkyl), —SO₂NH₂, —SO₂C₁₋₆ alkyl,—SO₂OC₁₋₆ alkyl, —OSO₂C₁₋₆ alkyl, —SOC₁₋₆ alkyl, —Si(C₁₋₆ alkyl)₃,—OSi(C₁₋₆ alkyl)₃-C(═S)N(C₁₋₆ alkyl)₂, C(═S)NH(C₁₋₆ alkyl), C(═S)NH₂,—C(═O)S(C₁₋₆ alkyl), —C(═S)SC₁₋₆ alkyl, —SC(═S)SC₁₋₆ alkyl, —P(═O)₂(C₁₋₆alkyl), —P(═O)(C₁₋₆ alkyl)₂, —OP(═O)(C₁₋₆ alkyl)₂, —OP(═O)(OC₁₋₆alkyl)₂, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₃₋₁₀ carbocyclyl, C₆₋₁₀ aryl, 3-10 membered heterocyclyl, 5-10 memberedheteroaryl; or two geminal R^(gg) substituents can be joined to form ═Oor ═S; wherein X⁻ is a counterion.

A “counterion” or “anionic counterion” is a negatively charged groupassociated with a cationic quaternary amino group in order to maintainelectronic neutrality. Exemplary counterions include halide ions (e.g.,F⁻, Cl⁻, Br⁻, I⁻), NO₃ ⁻, ClO₄ ⁻, OH⁻, H₂PO₄ ⁻, HSO₄ ⁻, sulfonate ions(e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate,benzenesulfonate, 10-camphor sulfonate, naphthalene-2-sulfonate,naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonicacid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate,ethanoate, propanoate, benzoate, glycerate, lactate, tartrate,glycolate, and the like).

“Halo” or “halogen” refers to fluorine (fluoro, —F), chlorine (chloro,—C₁), bromine (bromo, —Br), or iodine (iodo, —I).

Nitrogen atoms can be substituted or unsubstituted as valency permits,and include primary, secondary, tertiary, and quarternary nitrogenatoms. Exemplary nitrogen atom substitutents include, but are notlimited to, hydrogen, —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),—C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —C(═NR^(bb))R,—C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc),—SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc),—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂, —P(═O)(NR^(cc))₂,C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups attached to a nitrogen atom are joinedto form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5R^(dd) groups, and wherein R^(aa), R^(bb), R^(cc) and R^(dd) are asdefined above.

In certain embodiments, the substituent present on a nitrogen atom is anitrogen protecting group (also referred to as an amino protectinggroup). Nitrogen protecting groups include, but are not limited to, —OH,—OR^(aa), —N(R^(cc))₂, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(cc))R^(aa), —C(═NR^(cc))OR^(aa),—C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc), —SO₂OR^(cc),—SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc), C₁₋₁₀ alkyl(e.g., aralkyl, heteroaralkyl), C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl groups, wherein each alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aralkyl, aryl, and heteroaryl is independently substitutedwith 0, 1, 2, 3, 4, or 5 R^(dd) groups, and wherein R^(aa), R^(bb),R^(cc), and R^(dd) are as defined herein. Nitrogen protecting groups arewell known in the art and include those described in detail inProtecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts,3rd edition, John Wiley & Sons, 1999, incorporated herein by reference.

Amide nitrogen protecting groups (e.g., —C(═O)R^(aa)) include, but arenot limited to, formamide, acetamide, chloroacetamide,trichloroacetamide, trifluoroacetamide, phenylacetamide,3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide,N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide,o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide,(N′-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide,3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide,2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine,o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

Carbamate nitrogen protecting groups (e.g., —C(═O)OR^(aa)) include, butare not limited to, methyl carbamate, ethyl carbamante,9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate,2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methylcarbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate(Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate,1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC),1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC),1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2′- and4′-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethylcarbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinylcarbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate(Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc),8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate,2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate,2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc),2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate(Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc),1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate,p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate,2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate,3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methylcarbamate, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzylcarbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentylcarbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate,2,2-dimethoxyacylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate,isobutyl carbamate, isonicotinyl carbamate,p-(p′-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate,1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate,1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate,p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate,4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzylcarbamate.

Sulfonamide nitrogen protecting groups (e.g., —S(═O)₂R^(aa)) include,but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide,2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),2,4,6-trimethoxybenzenesulfonamide (Mtb),2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide(Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide(Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide,4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

Other nitrogen protecting groups include, but are not limited to,phenothiazinyl-(10)-acyl derivative, N′-p-toluenesulfonylaminoacylderivative, N′-phenylaminothioacyl derivative, N-benzoylphenylalanylderivative, N-acetylmethionine derivative,4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts),N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole,N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE),5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine,N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammoniumsalts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),N-9-phenylfluorenylamine (PhF),N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm),N-2-picolylamino N′-oxide, N-1,1-dimethylthiomethyleneamine,N-benzylideneamine, N-p-methoxybenzylideneamine,N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,N—(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine,N-p-nitrobenzylideneamine, N-salicylideneamine,N-5-chlorosalicylideneamine,N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,N-borane derivative, N-diphenylborinic acid derivative,N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate,N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide,diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt),diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzylphosphoramidate, diphenyl phosphoramidate, benzenesulfenamide,o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide,pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide,triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

In certain embodiments, the substituent present on an oxygen atom is anoxygen protecting group (also referred to as a hydroxyl protectinggroup). Oxygen protecting groups include, but are not limited to,—R^(aa), —N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa),—C(═O)N(R^(bb))₂, —C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa),—C(═NR^(bb))N(R^(bb))₂, —S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃,—P(R^(cc))₂, —P(R^(cc))₃, —P(═O)₂R^(aa), —P(═O)(R^(aa))₂,—P(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, and —P(═O)(NR^(bb))₂, whereinR^(aa), R^(bb), and R^(cc) are as defined herein. Oxygen protectinggroups are well known in the art and include those described in detailin Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M.Wuts, 3^(rd) edition, John Wiley & Sons, 1999, incorporated herein byreference.

Exemplary oxygen protecting groups include, but are not limited to,methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl,(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl,bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR),tetrahydropyranyl (THP), 3-bromotetrahydropyranyl,tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl(MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranylS,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl,t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl,benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl,p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido,diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl,triphenylmethyl, α-naphthyldiphenylmethyl,p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl,tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxyphenyl)diphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-tris(levulinoyloxyphenyl)methyl,4,4′,4″-tris(benzoyloxyphenyl)methyl,3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl,1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl,9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl(TMS), triethylsilyl (TES), triisopropylsilyl (TIPS),dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS),dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl(TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl,diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate,benzoylformate, acetate, chloroacetate, dichloroacetate,trichloroacetate, trifluoroacetate, methoxyacetate,triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate,3-phenylpropionate, 4-oxopentanoate (levulinate),4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate,adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate,2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate,9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate(TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec),2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutylcarbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkylp-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzylcarbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzylcarbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate,4-ethoxy-1-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate,4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,o-(methoxyacyl)benzoate, α-naphthoate, nitrate, alkylN,N,N′,N′-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate(Ts).

In certain embodiments, the substituent present on a sulfur atom is asulfur protecting group (also referred to as a thiol protecting group).Sulfur protecting groups include, but are not limited to, —R^(aa),—N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂,—S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃, —P(R^(cc))₂, —P(R^(cc))₃,—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, and—P(═O)(NR^(bb))₂, wherein R^(aa), R^(bb), and R^(cc) are as definedherein. Sulfur protecting groups are well known in the art and includethose described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P. G. M. Wuts, 3^(rd) edition, John Wiley & Sons, 1999,incorporated herein by reference.

These and other exemplary substituents are described in more detail inthe Detailed Description, Examples, and claims. The present disclosureis not intended to be limited in any manner by the above exemplarylisting of substituents.

“Pharmaceutically acceptable salt” refers to those salts which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and other animals without undue toxicity,irritation, allergic response, and the like, and are commensurate with areasonable benefit/risk ratio. Pharmaceutically acceptable salts arewell known in the art. For example, Berge et al. describepharmaceutically acceptable salts in detail in J. PharmaceuticalSciences (1977) 66:1-19. Pharmaceutically acceptable salts of thecompounds describe herein include those derived from suitable inorganicand organic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid, or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺ (C₁₋₄alkyl)₄ salts. Representativealkali or alkaline earth metal salts include sodium, lithium, potassium,calcium, magnesium, and the like. Further pharmaceutically acceptablesalts include, when appropriate, quaternary salts.

A “subject” to which administration is contemplated includes, but is notlimited to, humans (e.g., a male or female of any age group, e.g., apediatric subject (e.g, infant, child, adolescent) or adult subject(e.g., young adult, middle-aged adult or senior adult)) and/or othernon-human animals, for example, non-human mammals (e.g., primates (e.g.,cynomolgus monkeys, rhesus monkeys); commercially relevant mammals suchas cattle, pigs, horses, sheep, goats, cats, and/or dogs), birds (e.g.,commercially relevant birds such as chickens, ducks, geese, and/orturkeys), rodents (e.g., rats and/or mice), reptiles, amphibians, andfish. In certain embodiments, the non-human animal is a mammal. Thenon-human animal may be a male or female at any stage of development. Anon-human animal may be a transgenic animal.

“Condition,” “disease,” and “disorder” are used interchangeably herein.

“Treat,” “treating” and “treatment” encompasses an action that occurswhile a subject is suffering from a condition which reduces the severityof the condition or retards or slows the progression of the condition(“therapeutic treatment”). “Treat,” “treating” and “treatment” alsoencompasses an action that occurs before a subject begins to suffer fromthe condition and which inhibits or reduces the severity of thecondition (“prophylactic treatment”).

An “effective amount” of a compound refers to an amount sufficient toelicit the desired biological response, e.g., treat the condition. Aswill be appreciated by those of ordinary skill in this art, theeffective amount of a compound described herein may vary depending onsuch factors as the desired biological endpoint, the pharmacokinetics ofthe compound, the condition being treated, the mode of administration,and the age and health of the subject. An effective amount encompassestherapeutic and prophylactic treatment.

A “therapeutically effective amount” of a compound is an amountsufficient to provide a therapeutic benefit in the treatment of acondition or to delay or minimize one or more symptoms associated withthe condition. A therapeutically effective amount of a compound means anamount of therapeutic agent, alone or in combination with othertherapies, which provides a therapeutic benefit in the treatment of thecondition. The term “therapeutically effective amount” can encompass anamount that improves overall therapy, reduces or avoids symptoms orcauses of the condition, or enhances the therapeutic efficacy of anothertherapeutic agent.

A “prophylactically effective amount” of a compound is an amountsufficient to prevent a condition, or one or more symptoms associatedwith the condition or prevent its recurrence. A prophylacticallyeffective amount of a compound means an amount of a therapeutic agent,alone or in combination with other agents, which provides a prophylacticbenefit in the prevention of the condition. The term “prophylacticallyeffective amount” can encompass an amount that improves overallprophylaxis or enhances the prophylactic efficacy of anotherprophylactic agent.

As used herein, the term “methyltransferase” represents transferaseclass enzymes that are able to transfer a methyl group from a donormolecule to an acceptor molecule, e.g., an amino acid residue of aprotein or a nucleic base of a DNA molecule. Methytransferases typicallyuse a reactive methyl group bound to sulfur in S-adenosyl methionine(SAM) as the methyl donor. In some embodiments, a methyltransferasedescribed herein is a protein methyltransferase. In some embodiments, amethyltransferase described herein is a histone methyltransferase.Histone methyltransferases (HMT) are histone-modifying enzymes,(including histone-lysine N-methyltransferase and histone-arginineN-methyltransferase), that catalyze the transfer of one or more methylgroups to lysine and arginine residues of histone proteins. In certainembodiments, a methyltransferase described herein is a histone-arginineN-methyltransferase.

As generally described above, provided herein are compounds useful asPRMT5 inhibitors. In some embodiments, the present disclosure provides acompound of Formula (I):

or a pharmaceutically acceptable salt thereof,wherein

-   -   R¹ is hydrogen, R^(z), or —C(O)R^(z), wherein R^(z) is        optionally substituted C₁₋₆ alkyl;

L_(z) is a linker;

Ring Z is an optionally substituted, monocyclic or bicyclic, saturated,partially unsaturated, or aromatic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur;

R²¹, R²², R²³, and R²⁴ are independently hydrogen, halo, or optionallysubstituted aliphatic;

each R^(x) is independently selected from the group consisting of halo,—CN, optionally substituted aliphatic, and —OR′;

R′ is hydrogen or optionally substituted aliphatic; and

n is 0, 1, 2, 3, 4, 5, 6, 7, or 8.

In some embodiments, the present disclosure provides a compound ofFormula (I^(A)):

or a pharmaceutically acceptable salt thereof,wherein

R¹ is hydrogen, R^(z), or —C(O)R^(z), wherein R^(z) is optionallysubstituted C₁₋₆ alkyl;

X_(A) is a bond, —O—, —N(R)—, —CR^(4A)R^(5A)—, —O—CR^(4A)R^(5A),—N(R)—CR^(4A)R^(5A)—, —O—CR^(4A)R^(5A)—, —N(R)—CR^(4A)R^(5A)—O,—N(R)—CR^(4A)R^(5A)—N(R)—, —O—CR^(4A)R^(5A)—N(R)—, —CR^(4A)R^(5A)—O—,—CR^(4A)R^(5A)—N(R)—, —O—CR^(4A)R^(5A)—CR^(6A)R^(7A)—,—N(R)—CR^(4A)R^(5A)—CR^(6A)R^(7A)—, —CR^(6A)R^(7A)—CR^(4A)R^(5A)—O—,—CR^(6A)R^(7A)—CR^(4A)R^(5A)—N(R)—, or —CR^(6A)R^(7A)—CR^(4A)R^(5A)—;

each R is independently hydrogen or optionally substituted C₁₋₆aliphatic;

R^(2A) and R^(3A) are independently selected from the group consistingof hydrogen, halo, —CN, —NO₂, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted phenyl,optionally substituted heterocyclyl, optionally substituted heteroaryl,—OR^(A), N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or R^(2A)and R^(3A) are taken together with their intervening atoms to form anoptionally substituted carbocyclic or heterocyclic ring;

R^(4A) and R^(5A) are independently selected from the group consistingof hydrogen, halo, —CN, —NO₂, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted phenyl,optionally substituted heterocyclyl, optionally substituted heteroaryl,—OR^(A), N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or R^(4A)and R^(5A) are taken together with their intervening atoms to form anoptionally substituted carbocyclic or heterocyclic ring;

R^(6A) and R^(7A) are independently selected from the group consistingof hydrogen, halo, —CN, —NO₂, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted phenyl,optionally substituted heterocyclyl, optionally substituted heteroaryl,—OR^(A), N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or R^(6A)and R^(7A) are taken together with their intervening atoms to form anoptionally substituted carbocyclic or heterocyclic ring;

each R^(A) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl;

each R^(B) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl, or two R^(B) groups aretaken together with their intervening atoms to form an optionallysubstituted heterocyclic ring;

R^(8A), R^(9A), R^(10A), and R^(11A) are independently hydrogen, halo,or optionally substituted aliphatic;

Cy^(A) is a monocyclic or bicyclic, saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, wherein Cy^(A) is substituted with 0, 1,2, 3, or 4 R^(y) groups;

each R^(y) is independently selected from the group consisting of halo,—CN, —NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted aryl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂;

each R^(x) is independently selected from the group consisting of halo,—CN, optionally substituted aliphatic, and —OR′;

R′ is hydrogen or optionally substituted aliphatic; and

n is 0, 1, 2, 3, 4, 5, 6, 7, or 8.

In certain embodiments, a provided compound is of Formula (I^(A)-a):

or a pharmaceutically acceptable salt thereof, wherein R, R¹, R^(2A),R^(3A), R^(8A), R^(9A), R^(10A), R^(11A), R^(x) n, X_(A), and Cy^(A) areas described herein.

In certain embodiments, a provided compound is of Formula (I^(A)-b):

or a pharmaceutically acceptable salt thereof, wherein R, R¹, R^(2A),R^(3A), R^(8A), R^(9A), R^(10A), R^(11A), R^(x) n, X_(A), and Cy^(A) areas described herein.

In certain embodiments, a provided compound is of Formula (I^(A)-c):

or a pharmaceutically acceptable salt thereof, wherein R, R¹, R^(2A),R^(3A), R^(x), n, X_(A), and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (II^(A)):

or a pharmaceutically acceptable salt thereof, wherein R^(2A), R^(3A),R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (II^(A)-a):

or a pharmaceutically acceptable salt thereof, wherein R^(2A), R^(3A),R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (II^(A)-b):

or a pharmaceutically acceptable salt thereof, wherein R^(2A), R^(3A),R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (III^(A))

or a pharmaceutically acceptable salt thereof, wherein R, R^(2A),R^(3A), R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (III^(A)-a):

or a pharmaceutically acceptable salt thereof, wherein R, R^(2A),R^(3A), R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (III^(A)-b):

or a pharmaceutically acceptable salt thereof, wherein R, R^(2A),R^(3A), R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (IV^(A)):

or a pharmaceutically acceptable salt thereof, wherein R^(2A), R^(3A),R^(4A), R^(5A), R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (IV^(A)-a):

or a pharmaceutically acceptable salt thereof, wherein R^(2A), R^(3A),R^(4A), R^(5A), R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (IV^(A)-b):

or a pharmaceutically acceptable salt thereof, wherein R^(2A), R^(3A),R^(4A), R^(5A), R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (V^(A)):

or a pharmaceutically acceptable salt thereof, wherein R^(2A), R^(3A),R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (V^(A)-a):

or a pharmaceutically acceptable salt thereof, wherein R^(2A), R^(3A),R^(x), n, and Cy^(A) are as described herein.

In certain embodiments, a provided compound is of Formula (V^(A)-b):

or a pharmaceutically acceptable salt thereof, wherein R^(2A), R^(3A),R^(x), n, and Cy^(A) are as described herein.

In some embodiments, the present disclosure provides a compound ofFormula (I^(B)):

or a pharmaceutically acceptable salt thereof,wherein

R¹ is hydrogen, R^(z), or —C(O)R^(z), wherein R^(z) is optionallysubstituted C₁₋₆ alkyl;

L_(B) is —N(R)C(O)—, —C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)O—, or—OC(O)N(R)—;

each R is independently hydrogen or optionally substituted C₁₋₆aliphatic;

Ar is a monocyclic or bicyclic aromatic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, wherein Ar issubstituted with 0, 1, 2, 3, 4, or 5 R^(y) groups, as valency permits;

each R^(y) is independently selected from the group consisting of halo,—CN, —NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted aryl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂;

each R^(A) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl;

each R^(B) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl, or two R^(B) groups aretaken together with their intervening atoms to form an optionallysubstituted heterocyclic ring;

R^(5B), R^(6B), R^(7B), and R^(8B) are independently hydrogen, halo, oroptionally substituted aliphatic;

each R^(x) is independently selected from the group consisting of halo,—CN, optionally substituted aliphatic, and —OR′;

R′ is hydrogen or optionally substituted aliphatic; and

n is 0, 1, 2, 3, 4, 5, 6, 7, or 8.

In certain embodiments, a provided compound is of Formula (I^(B)-a):

or a pharmaceutically acceptable salt thereof, wherein R¹, R^(5B),R^(6B), R^(7B), R^(8B), R^(x), n, L_(B), and Ar are as described herein.

In certain embodiments, a provided compound is of Formula (I^(B)-b):

or a pharmaceutically acceptable salt thereof, wherein R¹, R^(5B),R^(6B), R^(7B), R^(8B), R^(x), n, L_(B), and Ar are as described herein.

In certain embodiments, a provided compound is of Formula (I^(B)-C):

or a pharmaceutically acceptable salt thereof, wherein R¹, R^(x), n,L_(B), and Ar are as described herein.

In certain embodiments, a provided compound is of Formula (II^(B)):

or a pharmaceutically acceptable salt thereof, wherein R¹, R^(x), n, andAr are as described herein.

In certain embodiments, a provided compound is of Formula (II^(B)-a):

or a pharmaceutically acceptable salt thereof, wherein R¹, R^(x), n, andAr are as described herein.

In certain embodiments, a provided compound is of Formula (II^(B)-b):

or a pharmaceutically acceptable salt thereof, wherein R¹, R^(x), n, andAr are as described herein.

In certain embodiments, a provided compound is of Formula (III^(B)):

or a pharmaceutically acceptable salt thereof, wherein R^(y), R^(x), andn are as described herein.

In certain embodiments, a provided compound is of Formula (III^(B)-a):

or a pharmaceutically acceptable salt thereof, wherein R^(y), R^(x), andn are as described herein.

In certain embodiments, a provided compound is of Formula (III^(B)-b):

or a pharmaceutically acceptable salt thereof, wherein R^(y), R^(x), andn are as described herein.

In some embodiments, the present disclosure provides a compound ofFormula (I^(C)):

or a pharmaceutically acceptable salt thereof;wherein

Ring C is an optionally substituted, 5- to 12-membered, monocyclic orbicyclic, heterocyclyl or heteroaryl having 1-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur;

R¹ is hydrogen, R^(z), or —C(O)R^(z), wherein R^(z) is optionallysubstituted C₁₋₆ alkyl;

Y is O or S;

R^(5B), R^(6B), R^(7B), and R^(8B) are independently hydrogen, halo, oroptionally substituted aliphatic;

each R^(x) is independently selected from the group consisting of halo,—CN, optionally substituted aliphatic, and —OR′;

R′ is hydrogen or optionally substituted aliphatic; and

n is 0, 1, 2, 3, 4, 5, 6, 7, or 8.

In certain embodiments, a provided compound is of Formula (I^(C)-a):

or a pharmaceutically acceptable salt thereof, wherein Ring C, Y, R¹,R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (I^(C)-b):

or a pharmaceutically acceptable salt thereof, wherein Ring C, Y, R¹,R^(5B), R^(6B), R^(7B), R^(8B), R^(x), andn are as described herein.

In certain embodiments, a provided compound is of Formula (I^(C)-c):

or a pharmaceutically acceptable salt thereof, wherein Ring C, Y, R¹,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (II^(C)):

or a pharmaceutically acceptable salt thereof, wherein R¹, R^(x), and nare as described herein,

G is NR^(2C), CR^(3C)R^(4C), O or S;

R^(2C) is selected from the group consisting of optionally substitutedaliphatic, optionally substituted carbocyclyl, optionally substitutedaryl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —C(O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂, —C(═S)R^(A), —C(═S)N(R^(B))₂,—S(═O)R^(A), —SO₂R^(A), and —SO₂N(R^(B))₂;

R^(3C) is selected from the group consisting of hydrogen, halo,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted aryl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂;

each R^(A) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl;

each R^(B) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl, or two R^(B) groups aretaken together with their intervening atoms to form an optionallysubstituted heterocyclic ring;

R^(4C) is selected from the group consisting of hydrogen, halo, oroptionally substituted aliphatic;

each R^(y) is independently selected from the group consisting of halo,—CN, —NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted aryl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂, or two adjacent R^(y) groups may betaken together with their intervening atoms to form a saturated,partially unsaturated, or aromatic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur;

p is 0, 1, or 2; and

k is 0, 1, 2, 3, or 4.

In certain embodiments, a provided compound is of Formula (II^(C)-a):

or a pharmaceutically acceptable salt thereof, wherein R¹, G, Y, R^(y),k, p, R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (II^(C)-b):

or a pharmaceutically acceptable salt thereof, wherein R¹, G, Y, R^(y),k, p, R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (III^(C)):

or a pharmaceutically acceptable salt thereof, wherein R^(2C), R^(y), k,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (III^(C)-a):

or a pharmaceutically acceptable salt thereof, wherein R^(2C), R^(y), k,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (III^(C)-b):

or a pharmaceutically acceptable salt thereof, wherein R^(2C), R^(y), k,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (IV^(C)):

or a pharmaceutically acceptable salt thereof, wherein R^(3C), R^(y), k,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (IV^(C)-a):

or a pharmaceutically acceptable salt thereof, wherein R^(3C), R^(y), k,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (IV^(C)-b):

or a pharmaceutically acceptable salt thereof, wherein R^(3C), R^(y), k,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (V^(C)):

or a pharmaceutically acceptable salt thereof, wherein R^(y), k, R^(x),and n are as described herein.

In certain embodiments, a provided compound is of Formula (V^(C)-a):

or a pharmaceutically acceptable salt thereof, wherein R^(y), k, R^(x),and n are as described herein.

In certain embodiments, a provided compound is of Formula (V^(C)-b):

or a pharmaceutically acceptable salt thereof, wherein R^(y), k, R^(x),and n are as described herein.

In certain embodiments, a provided compound is of Formula (VI^(C)):

or a pharmaceutically acceptable salt thereof, wherein R^(y), k, R^(x),and n are as described herein.

In certain embodiments, a provided compound is of Formula (VI^(C)-a):

or a pharmaceutically acceptable salt thereof, wherein R^(y), k, R^(x),and n are as described herein.

In certain embodiments, a provided compound is of Formula (VI^(C)-b)

or a pharmaceutically acceptable salt thereof, wherein R^(y), k, R^(x),and n are as described herein.

In some embodiments, the present disclosure provides a compound ofFormula (I^(D)):

or a pharmaceutically acceptable salt thereof,wherein

R¹ is hydrogen, R^(z), or —C(O)R^(z), wherein R^(z) is optionallysubstituted C₁₋₆ alkyl;

L_(D) is —O—, —N(R)—, —C(R^(2A))(R^(3A))—, —O—CR^(2A)R^(3A),—N(R)—CR^(2A)R^(3A)—, —O—CR^(2A)R^(3A)—, —N(R)—CR^(2A)R^(3A)—O,—N(R)—CR^(2A)R^(3A)—N(R)—, —O—CR^(2A)R^(3A)—N(R)—, —CR^(2A)R^(3A)—O—,—CR^(2A)R^(3A)—N(R)—, —O—CR^(2A)R^(3A)—CR⁹R¹⁰—,—N(R)—CR^(2A)R^(3A)—CR⁹R¹⁰—, —CR^(2A)R^(3A)—CR⁹R¹⁰—O—,—CR^(2A)R^(3A)—CR⁹R¹⁰—N(R)—, or —CR^(2A)R^(3A)—CR⁹R¹⁰—;

each R is independently hydrogen or optionally substituted C₁₋₆aliphatic;

R^(2A) and R^(3A) are independently selected from the group consistingof hydrogen, halo, —CN, —NO₂, optionally substituted aliphatic,optionally substituted carbocyclyl; optionally substituted phenyl,optionally substituted heterocyclyl, optionally substituted heteroaryl,—OR^(A), N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or R^(2A)and R^(3A) are taken together with their intervening atoms to form anoptionally substituted carbocyclic or heterocyclic ring;

each R^(A) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl;

each R^(B) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl, or two R^(B) groups aretaken together with their intervening atoms to form an optionallysubstituted heterocyclic ring;

Ring A is a monocyclic or bicyclic, saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur;

R⁴ is -L₁-Cy^(D);

L₁ is a bond, —O—, —S—, —N(R)—, —C(O)—, —C(O)N(R)—, —N(R)C(O)N(R)—,—N(R)C(O)—, —N(R)C(O)O—, —OC(O)N(R)—, —SO₂—, —SO₂N(R)—, —N(R)SO₂—,—OC(O)—, —C(O)O—, or an optionally substituted, straight or branched,C₁₋₆ aliphatic chain wherein one, two, or three methylene units of L₁are optionally and independently replaced by —O—, —S—, —N(R)—, —C(O)—,—C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)—, —N(R)C(O)O—, —OC(O)N(R)—, —SO₂—,—SO₂N(R)—, —N(R)SO₂—, —OC(O)—, or —C(O)O—;

Cy^(D) is an optionally substituted, monocyclic, bicyclic or tricyclic,saturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur;

R^(5B), R^(6B), R^(7B), and R^(8B) are independently hydrogen, halo, oroptionally substituted aliphatic;

R⁹ and R¹⁰ are independently selected from the group consisting ofhydrogen, halo, —CN, —NO₂, optionally substituted aliphatic, optionallysubstituted carbocyclyl; optionally substituted phenyl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or R⁹ andR¹⁰ are taken together with their intervening atoms to form anoptionally substituted carbocyclic or heterocyclic ring;

each R^(y) is independently selected from the group consisting of halo,—CN, —NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl; optionally substituted aryl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂;

each R^(x) is independently selected from the group consisting of halo,—CN, optionally substituted aliphatic, and —OR′;

R′ is hydrogen or optionally substituted aliphatic;

n is 0, 1, 2, 3, 4, 5, 6, 7, or 8;

m is 0, 1, 2, 3, 4, 5, 6, 7, or 8, as valency permits; and

q is 0 or 1.

In certain embodiments, a provided compound is of Formula (I^(D)-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L_(D),R¹, R⁴, R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, q, R^(x), and n are asdefined herein.

In certain embodiments, a provided compound is of Formula (I^(D)-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L_(D),R¹, R⁴, R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, q, R^(x), and n are asdefined herein.

In certain embodiments, a provided compound is of Formula (I^(D)-c):

or a pharmaceutically acceptable salt thereof, wherein Ring A, L_(D),R¹, R⁴, R, m, q, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (II^(D)):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R, R¹,R⁴, R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, q, R^(x), and n are asdefined herein.

In certain embodiments, a provided compound is of Formula (II^(D)-a)

or a pharmaceutically acceptable salt thereof, wherein Ring A, R, R¹,R⁴, R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, q, R^(x), and n are asdefined herein.

In certain embodiments, a provided compound is of Formula (II^(D)-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R, R¹,R⁴, R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, q, R^(x), and n are asdefined herein.

In certain embodiments, a provided compound is of Formula (II^(D)-c):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R, R¹,R⁴, R^(y), m, q, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (III^(D)):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹,R^(2A), R^(3A), R⁴, R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, q, R^(x),and n are as defined herein.

In certain embodiments, a provided compound is of Formula (III^(D)-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹,R^(2A), R^(3A), R⁴, R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, q, R^(x),and n are as defined herein.

In certain embodiments, a provided compound is of Formula (III^(D)-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹,R^(2A), R^(3A), R⁴, R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, q, R^(x),and n are as defined herein.

In certain embodiments, a provided compound is of Formula (III^(D)-C):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹,R^(2A), R^(3A), R⁴, R^(y), m, q, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IV^(D)):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R⁴,R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (IV^(D)-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R⁴,R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (IV^(D)-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R⁴,R^(5B), R^(6B), R^(7B), R^(8B), R^(y), m, R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (IV^(D)-C):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R¹, R⁴,R^(y), m, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (V^(D)):

or a pharmaceutically acceptable salt thereof, wherein X₁, X₂, X₃, X₄,L_(D), L₁, Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n areas defined herein.

In certain embodiments, a provided compound is of Formula (V^(D)-a):

or a pharmaceutically acceptable salt thereof, wherein X₁, X₂, X₃, X₄,L_(D), L₁, Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n areas defined herein.

In certain embodiments, a provided compound is of Formula (V^(D)-b):

or a pharmaceutically acceptable salt thereof, wherein X₁, X₂, X₃, X₄,L_(D), L₁, Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n areas defined herein.

In certain embodiments, a provided compound is of Formula (V^(D)):

or a pharmaceutically acceptable salt thereof, wherein X₁, X₂, X₃, X₄,L_(D), L₁, Cy^(D), R¹, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VI^(D)):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (VI^(D)-a):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (VI^(D)-b):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (VI^(D)-c):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VII^(D)):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (VII^(D)-a):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (VII^(D)-b):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (VII^(D)-c):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (VIII^(D)):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (VIII^(D)-a):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (VIII^(D)-b):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (VIII^(D)-c):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (IX^(D)):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x) and n are as definedherein.

In certain embodiments, a provided compound is of Formula (IX^(D)-a):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x) and n are as definedherein.

In certain embodiments, a provided compound is of Formula (IX^(D)-b):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (IX^(D)-c):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(x), and n are as defined herein.

In certain embodiments, a provided compound is of Formula (X^(D)):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (X^(D)-a):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (XD-b):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(5B), R^(6B), R^(7B), R^(8B), R^(x), and n are as definedherein.

In certain embodiments, a provided compound is of Formula (X^(D)-c):

or a pharmaceutically acceptable salt thereof, wherein L_(D), L₁,Cy^(D), R¹, R^(x), and n are as defined herein.

As defined generally above, R¹ is hydrogen, R^(z), or —C(O)R^(z),wherein R^(z) is optionally substituted C₁₋₆ alkyl. In certainembodiments, R¹ is hydrogen. In some embodiments, R¹ is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, R¹ is unsubstituted C₁₋₆alkyl. In certain embodiments, R¹ is methyl, ethyl, or propyl. In someembodiments, R¹ is —C(O)R^(z), wherein R^(z) is optionally substitutedC₁₋₆ alkyl. In certain embodiments, R¹ is —C(O)R^(z), wherein R^(z) isunsubstituted C₁₋₆ alkyl. In certain embodiments, R¹ is acetyl.

As defined generally above, L_(z) is a linker. In certain embodiments,L_(z) is —X_(A)—C(R^(2A))(R^(3A))C(═O)N(R)—, L_(B), or L_(D) asdescribed herein.

As defined generally above, Ring Z is an optionally substituted,monocyclic or bicyclic, saturated, partially unsaturated, or aromaticring having 0-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In certain embodiments, Ring Z is Ring A, Ring C,Cy^(A), or Ar as described herein.

As defined generally above, R²¹, R²², R²³, and R²⁴ are independentlyhydrogen, halo, or optionally substituted aliphatic. In someembodiments, R²¹, R²², R²³, and R²⁴ are hydrogen. In some embodiments,R²², R²³, and R²⁴ are hydrogen, and R²¹ is optionally substitutedaliphatic. In some embodiments, R²², R²³, and R²⁴ are hydrogen, and R²¹is optionally substituted C₁₋₆ aliphatic. In some embodiments, R²², R²³,and R²⁴ are hydrogen, and R²¹ is optionally substituted C₁₋₃ aliphatic.In some embodiments, R²², R²³, and R²⁴ are hydrogen, and R²¹ is methyl.In some embodiments, R²¹, R²², and R²³ are hydrogen, and R²⁴ isoptionally substituted aliphatic. In some embodiments, R²¹, R²², and R²³are hydrogen, and R²⁴ is optionally substituted C₁₋₆ aliphatic. In someembodiments, R²¹, R²², and R²³ are hydrogen, and R²⁴ is optionallysubstituted C₁₋₃ aliphatic. In some embodiments, R²¹, R²², and R²³ arehydrogen, and R²⁴ is methyl.

As defined generally above, X_(A) is a bond, —O—, —N(R)—,—CR^(4A)R^(5A)—, —O—CR^(4A)R^(5A), —N(R)—CR^(4A)R^(5A)—,—O—CR^(4A)R^(5A)—O—, —N(R)—CR^(4A)R^(5A)—O, —N(R)—CR^(4A)R^(5A)—N(R)—,—O—CR^(4A)R^(5A)—N(R)—, —CR^(4A)R^(5A)—O—, —CR^(4A)R^(5A)—N(R)—,—O—CR^(4A)R^(5A)—CR^(6A)R^(7A)—, —N(R)—CR^(4A)R^(5A)—CR^(6A)R^(7A)—,—CR^(6A)R^(7A)—CR^(4A)R^(5A)—O—, —CR^(6A)R^(7A)—CR^(4A)R^(5A)—N(R)—, or—CR^(6A)R^(7A)—CR^(4A)R^(5A)—. In certain embodiments, X_(A) is a bond,—O—, —N(R)—, or —CR⁴R⁵—, wherein R¹, R⁴, and R⁵ are as described herein.In certain embodiments, X_(A) is a bond. In certain embodiments, X_(A)is —O—. In some embodiments, X_(A) is —N(R)—. In certain embodiments,X_(A) is —NH—. In certain embodiments, X_(A) is —N(R)—, wherein R isoptionally substituted C₁₋₆ aliphatic. In certain embodiments, X_(A) is—N(R)—, wherein R is optionally substituted C₁₋₆ alkyl. In certainembodiments, X_(A) is —N(R)—, wherein R is unsubstituted C₁₋₆ alkyl. Incertain embodiments, X_(A) is —N(Me)-. In some embodiments, X_(A) is—CR^(4A)R^(5A)—. In certain embodiments, X_(A) is —CH₂—. In certainembodiments, X_(A) is —CH₂—O—.

As defined generally above, each R is independently hydrogen oroptionally substituted C₁₋₆ aliphatic. In certain embodiments, R ishydrogen. In some embodiments, R is optionally substituted C₁₋₆aliphatic. In some embodiments, R is substituted C₁₋₆ aliphatic. In someembodiments, R is unsubstituted C₁₋₆ aliphatic. In some embodiments, Ris optionally substituted C₁₋₆ alkyl. In some embodiments, R issubstituted C₁₋₆ alkyl. In some embodiments, R is unsubstituted C₁₋₆alkyl. In some embodiments, R is methyl, ethyl, or propyl.

As defined generally above, R^(2A) and R^(3A) are independently selectedfrom the group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or R^(2A) and R^(3A) are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring. In certain embodiments, R^(2A) andR^(3A) are independently selected from the group consisting of hydrogen,halo, —CN, —NO₂, optionally substituted aliphatic, optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —OC(O)R^(A), NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂,—NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A),—S(O)R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R^(2A) andR^(3A) are taken together with their intervening atoms to form anoptionally substituted carbocyclic or heterocyclic ring.

In certain embodiments, R^(2A) is hydrogen. In some embodiments, R^(2A)is not hydrogen. In some embodiments, R^(2A) is halo. In certainembodiments, R^(2A) is fluoro. In some embodiments, R^(2A) is optionallysubstituted aliphatic. In certain embodiments, R^(2A) is optionallysubstituted C₁₋₆ aliphatic. In certain embodiments, R^(2A) is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, R^(2A) is substitutedC₁₋₆ alkyl. In certain embodiments, R^(2A) is —CF₃, CHF₂, or CH₂F. Incertain embodiments, R^(2A) is unsubstituted C₁₋₆ alkyl. In certainembodiments, R^(2A) is methyl, ethyl, or propyl. In certain embodiments,R^(3A) is hydrogen. In some embodiments, R^(3A) is not hydrogen. In someembodiments, R^(3A) is halo. In certain embodiments, R^(3A) is fluoro.

In some embodiments, R^(3A) is optionally substituted aliphatic. Incertain embodiments, R³ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R^(3A) is optionally substituted C₁₋₆ alkyl. Incertain embodiments, R^(3A) is substituted C₁₋₆ alkyl. In certainembodiments, R^(3A) is —CF₃, CHF₂, or CH₂F. In certain embodiments,R^(3A) is unsubstituted C₁₋₆ alkyl. In certain embodiments, R^(3A) ismethyl, ethyl, or propyl. In some embodiments, R^(2A) and R^(3A) are thesame. In some embodiments, R^(2A) and R^(3A) are different. In someembodiments, R^(2A) and R^(3A) are each hydrogen. In some embodiments,R^(2A) is hydrogen and R^(3A) is not hydrogen. In some embodiments,R^(2A) is hydrogen and R^(3A) is optionally substituted aliphatic. Insome embodiments, R^(2A) is hydrogen and R^(3A) is C₁₋₆ alkyl. In someembodiments, R^(2A) is hydrogen and R^(3A) is methyl.

As defined generally above, R^(4A) and R^(5A) are independently selectedfrom the group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R^(4A) and R^(5A) are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring. In certain embodiments, R^(4A) andR^(5A) are independently selected from the group consisting of hydrogen,halo, —CN, —NO₂, optionally substituted aliphatic, optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂,—NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A),—S(O)R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R^(4A) andR^(5A) are taken together with their intervening atoms to form anoptionally substituted carbocyclic or heterocyclic ring.

In certain embodiments, R^(4A) is hydrogen. In some embodiments, R^(4A)is not hydrogen. In some embodiments, R^(4A) is halo. In certainembodiments, R^(4A) is fluoro. In some embodiments, R^(4A) is optionallysubstituted aliphatic. In certain embodiments, R^(4A) is optionallysubstituted C₁₋₆ aliphatic. In certain embodiments, R^(4A) is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, R^(4A) is substitutedC₁₋₆ alkyl. In certain embodiments, R^(4A) is —CF₃, CHF₂, or CH₂F. Incertain embodiments, R^(4A) is unsubstituted C₁₋₆ alkyl. In certainembodiments, R^(4A) is methyl, ethyl, or propyl. In certain embodiments,R^(5A) is hydrogen. In some embodiments, R^(5A) is not hydrogen. In someembodiments, R^(5A) is halo. In certain embodiments, R^(5A) is fluoro.

In some embodiments, R^(5A) is optionally substituted aliphatic. Incertain embodiments, R^(5A) is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R^(5A) is optionally substituted C₁₋₆ alkyl. Incertain embodiments, R^(5A) is substituted C₁₋₆ alkyl. In certainembodiments, R^(5A) is —CF₃, CHF₂, or CH₂F. In certain embodiments,R^(5A) is unsubstituted C₁₋₆ alkyl. In certain embodiments, R^(5A) ismethyl, ethyl, or propyl. In some embodiments, R^(4A) and R^(5A) are thesame. In some embodiments, R^(4A) and R^(5A) are different. In someembodiments, R^(4A) and R^(5A) are each hydrogen. In some embodiments,R^(4A) is hydrogen and R^(5A) is not hydrogen. In some embodiments,R^(4A) is hydrogen and R^(5A) is optionally substituted aliphatic. Insome embodiments, R^(4A) is hydrogen and R^(5A) is C₁₋₆ alkyl. In someembodiments, R^(4A) is hydrogen and R^(5A) is methyl.

As defined generally above, R^(6A) and R^(7A) are independently selectedfrom the group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or R^(6A) and R^(7A) are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring. In certain embodiments, R^(6A) andR^(7A) are independently selected from the group consisting of hydrogen,halo, —CN, —NO₂, optionally substituted aliphatic, optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —OC(O)R^(A), NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂,—NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A),—S(O)R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R^(6A) andR^(7A) are taken together with their intervening atoms to form anoptionally substituted carbocyclic or heterocyclic ring.

In certain embodiments, R^(6A) is hydrogen. In some embodiments, R^(6A)is not hydrogen. In some embodiments, R^(6A) is halo. In certainembodiments, R^(6A) is fluoro. In some embodiments, R^(6A) is optionallysubstituted aliphatic. In certain embodiments, R^(6A) is optionallysubstituted C₁₋₆ aliphatic. In certain embodiments, R^(6A) is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, R^(6A) is substitutedC₁₋₆ alkyl. In certain embodiments, R^(6A) is —CF₃, CHF₂, or CH₂F. Incertain embodiments, R^(6A) is unsubstituted C₁₋₆ alkyl. In certainembodiments, R^(6A) is methyl, ethyl, or propyl. In certain embodiments,R^(7A) is hydrogen. In some embodiments, R^(7A) is not hydrogen. In someembodiments, R^(7A) is halo. In certain embodiments, R^(7A) is fluoro.

In some embodiments, R^(7A) is optionally substituted aliphatic. Incertain embodiments, R^(7A) is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R^(7A) is optionally substituted C₁₋₆ alkyl. Incertain embodiments, R^(7A) is substituted C₁₋₆ alkyl. In certainembodiments, R^(7A) is —CF₃, CHF₂, or CH₂F. In certain embodiments,R^(7A) is unsubstituted C₁₋₆ alkyl. In certain embodiments, R^(7A) ismethyl, ethyl, or propyl. In some embodiments, R^(6A) and R^(7A) are thesame. In some embodiments, R^(6A) and R^(7A) are different. In someembodiments, R^(6A) and R^(7A) are each hydrogen. In some embodiments,R^(6A) is hydrogen and R^(7A) is not hydrogen. In some embodiments,R^(6A) is hydrogen and R^(7A) is optionally substituted aliphatic. Insome embodiments, R^(6A) is hydrogen and R^(7A) is C₁₋₆ alkyl. In someembodiments, R^(6A) is hydrogen and R^(7A) is methyl.

As defined generally above, R^(8A), R^(9A), R^(10A), and R^(11A) areindependently hydrogen, halo, or optionally substituted aliphatic. Insome embodiments, R^(8A), R^(9A), R^(10A), and R^(11A) are hydrogen. Insome embodiments, R^(9A), R^(10A), and R^(11A) are hydrogen, and R^(8A)is optionally substituted aliphatic. In some embodiments, R^(9A),R^(10A), and R^(11A) are hydrogen, and R^(8A) is optionally substitutedC₁₋₆ aliphatic. In some embodiments, R^(9A), R^(10A), and R^(11A) arehydrogen, and R^(8A) is optionally substituted C₁₋₃ aliphatic. In someembodiments, R^(9A), R^(10A), and R^(11A) are hydrogen, and R^(8A) ismethyl. In some embodiments, R^(8A), R^(9A), and R^(10A) are hydrogen,and R^(11A) is optionally substituted aliphatic. In some embodiments,R^(8A), R^(9A), and R^(10A) are hydrogen, and R^(11A) is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R^(8A), R^(9A), andR^(10A) are hydrogen, and R^(11A) is optionally substituted C₁₋₃aliphatic. In some embodiments, R^(8A), R^(9A), and R^(10A) arehydrogen, and R^(11A) is methyl.

As defined generally above, R^(5B), R^(6B), R^(7B), and R^(8B) areindependently hydrogen, halo, or optionally substituted aliphatic. Insome embodiments, R^(5B), R^(6B), R^(7B), and R^(8B) are hydrogen. Insome embodiments, R^(6B), R^(7B), and R^(8B) are hydrogen, and R^(5B) isoptionally substituted aliphatic. In some embodiments, R^(6B), R^(7B),and R^(8B) are hydrogen, and R^(5B) is optionally substituted C₁₋₆aliphatic. In some embodiments, R^(6B), R^(7B), and R^(8B) are hydrogen,and R^(5B) is optionally substituted C₁₋₃ aliphatic. In someembodiments, R^(6B), R^(7B), and R^(8B) are hydrogen, and R^(5B) ismethyl. In some embodiments, R^(5B), R^(6B), and R^(7B) are hydrogen,and R^(8B) is optionally substituted aliphatic. In some embodiments,R^(5B), R^(6B), and R^(7B) are hydrogen, and R^(8B) is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R^(5B), R^(6B), andR^(7B) are hydrogen, and R^(8B) is optionally substituted C₁₋₃aliphatic. In some embodiments, R^(5B), R^(6B), and R^(7B) are hydrogen,and R^(8B) is methyl.

As defined generally above, L_(B) is —N(R)C(O)—, —C(O)N(R)—,—N(R)C(O)N(R)—, —N(R)C(O)O—, or —OC(O)N(R)—, wherein R is as describedherein. In some embodiments, L_(B) is —N(R)C(O)—. In some embodiments,L_(B) is —NHC(O)—. In some embodiments, L_(B) is —N(C₁₋₆ alkyl)C(O)—. Insome embodiments, L_(B) is —N(CH₃)C(O)—. In some embodiments, L_(B) is—C(O)N(R)—. In some embodiments, L_(B) is —C(O)NH—. In some embodiments,L_(B) is —C(O)N(C₁₋₆ alkyl)-. In some embodiments, L_(B) is—C(O)N(CH₃)—. In some embodiments, L_(B) is —N(R)C(O)N(R)—. In someembodiments, L_(B) is —NHC(O)NH—. In some embodiments, L_(B) is—NHC(O)N(R)—. In some embodiments, L_(B) is —N(R)C(O)NH—. In someembodiments, L_(B) is —N(CH₃)C(O)N(R)—. In some embodiments, L_(B) is—N(R)C(O)N(CH₃)—. In some embodiments, L_(B) is —N(CH₃)C(O)N(CH₃)—. Insome embodiments, L_(B) is —N(R)C(O)O—. In some embodiments, L_(B) is—NHC(O)O—. In some embodiments, L_(B) is —N(C₁₋₆ alkyl)C(O)O—. In someembodiments, L_(B) is —N(CH₃)C(O)O—. In some embodiments, L_(B) is—OC(O)N(R)—. In some embodiments, L_(B) is —OC(O)NH—. In someembodiments, L_(B) is —OC(O)N(C₁₋₆ alkyl)-. In some embodiments, L_(B)is —OC(O)N(CH₃)—.

As defined generally above, Ar is a monocyclic or bicyclic aromatic ringhaving 0-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein Ar is substituted with 0, 1, 2, 3, 4, or 5 R^(y) groups,as valency permits. In certain embodiments, Ar is unsubstituted. Incertain embodiments, Ar is substituted with one or two R^(y) groups. Incertain embodiments, Ar is substituted with one R^(y) group. In certainembodiments, Ar is substituted with two R^(y) groups. In certainembodiments, Ar is substituted with three R^(y) groups. In certainembodiments, Ar is substituted with four R^(y) groups. In certainembodiments, Ar is substituted with five R^(y) groups.

In certain embodiments, Ar is phenyl substituted with 0, 1, 2, 3, 4, or5 R^(y) groups. In certain embodiments, Ar is phenyl substituted withone or two R^(y) groups. In certain embodiments, Ar is unsubstitutedphenyl. In certain embodiments, Ar is phenyl substituted with one R^(y)group. In certain embodiments, Ar is phenyl substituted with two R^(y)groups. In certain embodiments, Ar is phenyl substituted with threeR^(y) groups. In certain embodiments, Ar is phenyl substituted with fourR^(y) groups. In certain embodiments, Ar is phenyl substituted with fiveR^(y) groups.

In certain embodiments, Ar is heteroaryl substituted with 0, 1, 2, 3, 4,or 5 R^(y) groups, as valency permits. In certain embodiments, Ar is a5- to 6-membered heteroaryl having 1-3 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur, and is substituted with 0,1, 2, 3, or 4 R^(y) groups. In certain embodiments, Ar is anunsubstituted 5- to 6-membered heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In certainembodiments, Ar is a 5- to 6-membered heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, and issubstituted with one or two R^(y) groups. In certain embodiments, Ar isa 5- to 6-membered heteroaryl having 1-3 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur, and is substituted with oneR^(y) group. In certain embodiments, Ar is a 5-membered heteroarylhaving 1-3 heteroatoms independently selected from nitrogen, oxygen, andsulfur (e.g., furanyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl,thiazolyl, imidazolyl, pyrazolyl, isothiazolyl, triazolyl, oxadiazolyl,thiadiazolyl), and is substituted with 0, 1, 2, 3, or 4 R^(y) groups. Incertain embodiments, Ar is a 6-membered heteroaryl having 1-3 nitrogens(e.g., pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, triazinyl), and issubstituted with 0, 1, 2, 3, 4, or 5 R^(y) groups.

In certain embodiments, Ar is a bicyclic aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein Ar is substituted with 0, 1, 2, 3, or 4 R^(y) groups. In certainembodiments, Ar is an 8- to 12-membered bicyclic aromatic ring having0-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein Ar is substituted with 0, 1, 2, 3, or 4 R^(y) groups. Incertain embodiments, Ar is an unsubstituted bicyclic aromatic ringhaving 0-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, Ar is a bicyclic aromatic ring having0-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein Ar is substituted with one or two R^(y) groups. Incertain embodiments, Ar is a bicyclic aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein Ar is substituted with one R^(y) group. In certain embodiments,Ar is a bicyclic aromatic ring having 0-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur, wherein Ar is substitutedwith two R^(y) groups. In certain embodiments, Ar is a bicyclic aromaticring having 0-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur, wherein Ar is substituted with three R^(y) groups.In certain embodiments, Ar is a bicyclic aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein Ar is substituted with four R^(y) groups. In certainembodiments, Ar is a bicyclic aromatic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, wherein Ar issubstituted with five R^(y) groups. In certain embodiments, Ar isnaphthalene substituted with 0, 1, 2, 3, 4, or 5 R^(y) groups.

In certain embodiments, Ar is an 8- to 10-membered bicyclic heteroarylhaving 1-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein Ar is substituted with 0, 1, 2, 3, or 4 R^(y) groups. Incertain embodiments, Ar is a 9-membered bicyclic heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur(e.g., indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl,isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl,benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl,benzisothiazolyl, benzthiadiazolyl, indolizinyl), wherein Ar issubstituted with 0, 1, 2, 3, 4, or 5 R^(y) groups. In certainembodiments, Ar is a 10-membered bicyclic heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur(e.g., naphthyridinyl, quinolinyl, isoquinolinyl, quinoxalinyl,quinazolinyl), wherein Ar is substituted with 0, 1, 2, 3, 4, or 5 R^(y)groups. In certain embodiments, Ar is selected from the group consistingof quinoline, benzimidazole, benzopyrazole, quinoxaline,tetrahydroquinoline, tetrahydroisoquinoline, naphthalene,tetrahydronaphthalene, 2,3-dihydrobenzo[b][1,4]dioxine, isoindole,2H-benzo[b][1,4]oxazin-3(4H)-one, 3,4-dihydro-2H-benzo[b][1,4]oxazine,and quinoxalin-2(1H)-one, wherein Ar is substituted with 0, 1, 2, 3, or4 R^(y) groups.

As defined generally above, Cy^(A) is a monocyclic or bicyclic,saturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein Cy^(A) is substituted with 0, 1, 2, 3, or 4 R^(y) groups. Incertain embodiments, Cy^(A) is unsubstituted. In certain embodiments,Cy^(A) is substituted with one or two R^(y) groups. In certainembodiments, Cy^(A) is substituted with one R^(y) group. In certainembodiments, Cy^(A) is substituted with two R^(y) groups. In certainembodiments, Cy^(A) is substituted with three R^(y) groups. In certainembodiments, Cy^(A) is substituted with four R^(y) groups.

In certain embodiments, Cy^(A) is phenyl substituted with 0, 1, 2, 3, or4 R^(y) groups. In certain embodiments, Cy^(A) is phenyl substitutedwith one or two R^(y) groups. In certain embodiments, Cy^(A) isunsubstituted phenyl. In certain embodiments, Cy^(A) is phenylsubstituted with one R^(y) group. In certain embodiments, Cy^(A) isphenyl substituted with two R^(y) groups. In certain embodiments, Cy^(A)is phenyl substituted with three R^(y) groups. In certain embodiments,Cy^(A) is phenyl substituted with four R^(y) groups.

In certain embodiments, Cy^(A) is a 5- to 6-membered heteroaryl having1-3 heteroatoms independently selected from nitrogen, oxygen, andsulfur, and is substituted with 0, 1, 2, 3, or 4 R^(y) groups. Incertain embodiments, Cy^(A) is an unsubstituted 5- to 6-memberedheteroaryl having 1-3 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In certain embodiments, Cy^(A) is a 5- to 6-memberedheteroaryl having 1-3 heteroatoms independently selected from nitrogen,oxygen, and sulfur, and is substituted with one or two R^(y) groups. Incertain embodiments, Cy^(A) is a 5- to 6-membered heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur,and is substituted with one R^(y) group. In certain embodiments, Cy^(A)is a 5-membered heteroaryl having 1-3 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur (e.g., furanyl, thienyl, pyrrolyl,oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyrazolyl, isothiazolyl,triazolyl, oxadiazolyl, thiadiazolyl), and is substituted with 0, 1, 2,3, or 4 R^(y) groups. In certain embodiments, Cy^(A) is a 6-memberedheteroaryl having 1-3 nitrogens (e.g., pyridyl, pyrimidyl, pyridazinyl,pyrazinyl, triazinyl), and is substituted with 0, 1, 2, 3, or 4 R^(y)groups.

In certain embodiments, Cy^(A) is a bicyclic saturated, partiallyunsaturated, or aromatic ring having 0-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur, wherein Cy^(A) issubstituted with 0, 1, 2, 3, or 4 R^(y) groups. In certain embodiments,Cy^(A) is an 8- to 12-membered bicyclic saturated, partiallyunsaturated, or aromatic ring having 0-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur, wherein Cy^(A) issubstituted with 0, 1, 2, 3, or 4 R^(y) groups. In certain embodiments,Cy^(A) is an unsubstituted bicyclic saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, Cy^(A) is abicyclic saturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein Cy^(A) is substituted with one or two R^(y) groups. In certainembodiments, Cy^(A) is a bicyclic saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, wherein Cy^(A) is substituted with oneR^(y) group. In certain embodiments, Cy^(A) is a bicyclic saturated,partially unsaturated, or aromatic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, wherein Cy issubstituted with two R^(y) groups. In certain embodiments, Cy^(A) is abicyclic saturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein Cy^(A) is substituted with three R^(y) groups. In certainembodiments, Cy^(A) is a bicyclic saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, wherein Cy^(A) is substituted with fourR^(y) groups.

In certain embodiments, Cy^(A) is an 8- to 10-membered bicyclicheteroaryl having 1-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur, wherein Cy^(A) is substituted with 0, 1, 2, 3, or 4R^(y) groups. In certain embodiments, Cy^(A) is a 9-membered bicyclicheteroaryl having 1-3 heteroatoms independently selected from nitrogen,oxygen, and sulfur (e.g., indolyl, isoindolyl, indazolyl,benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl,benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl,benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl,indolizinyl), wherein Cy^(A) is substituted with 0, 1, 2, 3, or 4 R^(y)groups. In certain embodiments, Cy^(A) is a 10-membered bicyclicheteroaryl having 1-3 heteroatoms independently selected from nitrogen,oxygen, and sulfur (e.g., naphthyridinyl, quinolinyl, isoquinolinyl,quinoxalinyl, quinazolinyl), wherein Cy^(A) is substituted with 0, 1, 2,3, or 4 R^(y) groups. In certain embodiments, Cy^(A) is selected fromthe group consisting of quinoline, benzimidazole, benzopyrazole,quinoxaline, tetrahydroquinoline, tetrahydroisoquinoline, naphthalene,tetrahydronaphthalene, 2,3-dihydrobenzo[b][1,4]dioxine, isoindole,2H-benzo[b][1,4]oxazin-3 (4H)-one, 3,4-dihydro-2H-benzo[b][1,4]oxazine,and quinoxalin-2(1H)-one, wherein Cy^(A) is substituted with 0, 1, 2, 3,or 4 R^(y) groups.

As defined generally above, each R^(y) is independently selected fromthe group consisting of halo, —CN, —NO₂, optionally substitutedaliphatic, optionally substituted carbocyclyl, optionally substitutedphenyl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A),—OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂, wherein R^(A) and R^(B) aredescribed herein. In certain embodiments, each R^(y) is independentlyselected from the group consisting of halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂, wherein R^(A) and R^(B) aredescribed herein.

In some embodiments, at least one R^(y) is halo. In certain embodiments,at least one R^(y) is fluoro. In certain embodiments, at least one R^(y)is chloro. In some embodiments, at least one R^(y) is —CN. In someembodiments, at least one R^(y) is —OR^(A), wherein R^(A) is optionallysubstituted aliphatic. In some embodiments, at least one R^(y) is—OR^(A), wherein R^(A) is unsubstituted C₁₋₆ alkyl. In certainembodiments, at least one R^(y) is methoxy, ethoxy, or propoxy. Incertain embodiments, at least one R^(y) is methoxy. In some embodiments,at least one R^(y) is —OR^(A), wherein R^(A) is substituted C₁₋₆ alkyl.In certain embodiments, at least one R^(y) is —OCH₂CH₂N(CH₃)₂. In someembodiments, at least one R^(y) is —N(R^(B))₂. In some embodiments, atleast one R^(y) is —N(R^(B))₂, wherein each R^(B) is independentlyselected from hydrogen or C₁₋₆ alkyl. In some embodiments, at least oneR^(y) is —NHR^(B). In some embodiments, at least one R^(y) is —N(C₁₋₆alkyl)₂, —NH(C₁₋₆ alkyl), or —NH₂. In certain embodiments, at least oneR^(y) is —NH₂. In certain embodiments, at least one R^(y) is —NHCH₃. Incertain embodiments, at least one R^(y) is —N(CH₃)₂.

In some embodiments, at least one R^(y) is optionally substitutedaliphatic. In certain embodiments, at least one R^(y) is substitutedaliphatic. In certain embodiments, at least one R^(y) is unsubstitutedaliphatic. In some embodiments, at least one R^(y) is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) isunsubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) issubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) ismethyl, ethyl, or propyl. In certain embodiments, at least one R^(y) ismethyl. In certain embodiments, at least one R^(y) is —CF₃, CHF₂, orCH₂F. In certain embodiments, at least one R^(y) is C₁₋₆ alkylsubstituted with aryl, heteroaryl, or heterocyclyl. In certainembodiments, at least one R^(y) is benzyl. In certain embodiments, atleast one R^(y) is —(C₁₋₆ alkyl)-heteroaryl. In certain embodiments, atleast one R^(y) is —(C₁₋₆ alkyl)-heterocyclyl. In certain embodiments,at least one R^(y) is —CH₂-heteroaryl. In certain embodiments, at leastone R^(y) is —CH₂-heterocyclyl.

In some embodiments, at least one R^(y) is —C(O)N(R^(B))₂. In certainembodiments, at least one R^(y) is —C(O)NHR^(B). In certain embodiments,at least one R^(y) is —C(O)NH₂. In certain embodiments, at least oneR^(y) is —C(O)N(R^(B))₂, wherein the R^(B) groups are taken togetherwith their intervening atoms to form an optionally substituted 5- to6-membered heterocyclyl. In certain embodiments, at least one R^(y) is—C(O)N(R^(B))₂, wherein the R^(B) groups are taken together with theirintervening atoms to form an optionally substituted morpholinyl.

In some embodiments, at least one R^(y) is —SO₂N(R^(B))₂. In certainembodiments, at least one R^(y) is —SO₂NHR^(B). In certain embodiments,at least one R^(y) is —SO₂NH₂. In certain embodiments, at least oneR^(y) is —SO₂N(R^(B))₂, wherein neither R^(B) is hydrogen. In certainembodiments, at least one R^(y) is —SO₂NH(C₁₋₆ alkyl) or —SO₂N(C₁₋₆alkyl)₂. In certain embodiments, at least one R^(y) is —SO₂N(CH₃)₂. Incertain embodiments, at least one R^(y) is —SO₂N(R^(B))₂, wherein theR^(B) groups are taken together with their intervening atoms to form anoptionally substituted 5- to 6-membered heterocyclyl. In certainembodiments, at least one R^(y) is —SO₂-morpholinyl. In certainembodiments, at least one R^(y) is —SO₂-piperidinyl, —SO₂-piperazinyl,or —SO₂-piperidinyl.

In some embodiments, at least one R^(y) is —SO₂R^(A). In someembodiments, at least one R^(y) is —SO₂R^(A), wherein R^(A) isoptionally substituted aliphatic. In some embodiments, at least oneR^(y) is —SO₂(C₁₋₆ alkyl). In some embodiments, at least one R^(y) is—SO₂CH₃. In some embodiments, at least one R^(y) is —C(O)R^(A). In someembodiments, at least one R^(y) is —C(O)R^(A), wherein R^(A) isoptionally substituted aliphatic. In some embodiments, at least oneR^(y) is —C(O)(C₁₋₆ alkyl). In some embodiments, at least one R^(y) is—C(O)CH₃.

In some embodiments, at least one R^(y) is —N(R^(B))C(O)R^(A). Incertain embodiments, at least one R^(y) is —NHC(O)R^(A). In certainembodiments, at least one R^(y) is —NHC(O)(C₁₋₆ alkyl).

In certain embodiments, at least one R^(y) is —NHC(O)CH₃.

In some embodiments, at least one R^(y) is —N(R^(B))SO₂R^(A). In someembodiments, at least one R^(y) is —NHSO₂R^(A). In some embodiments, atleast one R^(y) is —N(C₁₋₆ alkyl)SO₂R^(A). In certain embodiments, atleast one R^(y) is —NHSO₂(C₁₋₆ alkyl) or —N(C₁₋₆ alkyl)SO₂(C₁₋₆ alkyl).In certain embodiments, at least one R^(y) is —NHSO₂CH₃. In certainembodiments, at least one R^(y) is —N(CH₃)SO₂CH₃.

In some embodiments, at least one R^(y) is optionally substitutedheterocyclyl, optionally substituted carbocyclyl, optionally substitutedaryl, or optionally substituted heteroaryl. In certain embodiments, atleast one R^(y) is an optionally substituted 5- to 6-memberedheterocyclyl having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, at least one R^(y)is an optionally substituted 5-membered heterocyclyl having oneheteroatom selected from nitrogen, oxygen, and sulfur. In certainembodiments, at least one R^(y) is optionally substituted pyrrolidinyl.In certain embodiments, at least one R^(y) is pyrroldinyl,hydroxypyrrolidinyl, or methylpyrrolidinyl. In certain embodiments, atleast one R^(y) is an optionally substituted 6-membered heterocyclylhaving 1-2 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, at least one R^(y) is an optionallysubstituted 6-membered heterocyclyl having one heteroatom selected fromnitrogen, oxygen, and sulfur. In certain embodiments, at least one R^(y)is optionally substituted piperidinyl. In certain embodiments, at leastone R^(y) is an optionally substituted 6-membered heterocyclyl havingtwo heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, at least one R^(y) is optionallysubstituted piperdinyl, optionally substituted piperazinyl, oroptionally substituted morpholinyl. In certain embodiments, at least oneR^(y) is morpholinyl, tetrahydropyranyl, piperidinyl, methylpiperidinyl,piperazinyl, methylpiperazinyl, acetylpiperazinyl,methylsulfonylpiperazinyl, aziridinyl, or methylaziridinyl. In someembodiments, at least one R^(y) is an optionally substituted 5- to6-membered heteroaryl having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, at least one R^(y)is an optionally substituted 5-membered heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur. Incertain embodiments, at least one R^(y) is an optionally substituted5-membered heteroaryl having one heteroatom selected from nitrogen,oxygen, and sulfur. In certain embodiments, at least one R^(y) is anoptionally substituted 5-membered heteroaryl having two heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In certainembodiments, at least one R^(y) is an optionally substituted 6-memberedheteroaryl having 1-3 nitrogens. In certain embodiments, at least oneR^(y) is an optionally substituted pyrazolyl. In certain embodiments, atleast one R^(y) is an optionally substituted imidazolyl. In certainembodiments, at least one R^(y) is an optionally substituted pyridyl. Incertain embodiments, at least one R^(y) is an optionally substitutedpyrimidyl. In certain embodiments, at least one R^(y) is pyrazolyl,methylpyrazolyl, imidazolyl, or methylimidazolyl.

In some embodiments, for compounds of formula (II^(C)), (II^(C)-a),(II^(C)b), (III^(C)), (III^(C)-a), (III^(C)-b), (IV^(C)), (IV^(C)-a),(IV^(C)-b), (V^(C)), (V^(C)-a), (V^(C)-b), (VI^(C)), (VI^(C)-a), or(VI^(C)-b), two adjacent R^(y) groups may be taken together with theirintervening atoms to form a saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, two adjacent R^(y)groups may be taken together with their intervening atoms to form asaturated carbocyclic ring. In some embodiments, two adjacent R^(y)groups may be taken together with their intervening atoms to form apartially unsaturated carbocyclic ring. In some embodiments, twoadjacent R^(y) groups may be taken together with their intervening atomsto form a benzene ring. In some embodiments, two adjacent R^(y) groupsmay be taken together with their intervening atoms to form a saturatedring having 1-2 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In some embodiments, two adjacent R^(y) groups maybe taken together with their intervening atoms to form a partiallyunsaturated ring having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, two adjacent R^(y)groups may be taken together with their intervening atoms to form anaromatic ring having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur.

As defined generally above, Ring C is an optionally substituted, 5- to12-membered, monocyclic or bicyclic, heterocyclyl or heteroaryl having1-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur. One of ordinary skill in the art will understand that Ring Ccomprises an amide or thioamide. In certain embodiments, Ring C is anoptionally substituted, 5- to 6-membered, monocyclic heteroaryl having1-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, Ring C is an optionally substituted, 5-to 7-membered, monocyclic heterocyclyl having 1-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In certainembodiments, Ring C is an optionally substituted, 8- to 10-membered,bicyclic heteroaryl having 1-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, Ring C is anoptionally substituted, 8- to 12-membered, bicyclic heterocyclyl having1-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, Ring C is an optionally substitutedpiperdinone. In certain embodiments, Ring C is an optionally substitutedpyridinone. In certain embodiments, Ring C is an optionally substitutedpiperazinone. In certain embodiments, Ring C is an optionallysubstituted isoindolinone. In certain embodiments, Ring C is anoptionally substituted 2H-benzo[b][1,4]oxazin-3(4H)-one. In someembodiments, Ring C is

wherein G, R^(y), m, and p are as described herein.

In certain embodiments, Y is O. In certain embodiments, Y is S.

As defined generally above, G is NR^(2C), CR^(3C)R^(4C), O or S. Incertain embodiments, G is NR^(2C). In certain embodiments, G isCR^(3C)R^(4C). In certain embodiments, G is O. In certain embodiments, Gis S.

As defined generally above, R^(2C) is selected from the group consistingof optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted aryl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —C(O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂, —C(═S)R^(A),—C(═S)N(R^(B))₂, —S(═O)R^(A), —SO₂R^(A), and —SO₂N(R^(B))₂. In someembodiments, R^(2C) is optionally substituted aryl. In certainembodiments, R^(2C) is optionally substituted phenyl. In certainembodiments, R^(2C) is unsubstituted phenyl. In certain embodiments,R^(2C) is halophenyl. In certain embodiments, R^(2C) is fluorophenyl. Incertain embodiments, R^(2C) is chlorophenyl. In some embodiments, R^(2C)is phenyl substituted with optionally substituted C₁₋₆ alkyl. In someembodiments, R^(2C) is phenyl substituted with optionally substitutedC₁₋₃ alkyl. In certain embodiments, R^(2C) is phenyl substituted withmethyl. In certain embodiments, R^(2C) is phenyl substituted with—CH₂OH. In some embodiments, R^(2C) is phenyl substituted with aheterocyclic ring. In certain embodiments, R^(2C) is phenyl substitutedwith morpholinyl. In certain embodiments, R^(2C) is phenyl substitutedwith tetrahydropyranyl. In some embodiments, R^(2C) is optionallysubstituted heteroaryl. In certain embodiments, R^(2C) is optionallysubstituted quinoline. In certain embodiments, R^(2C) is unsubstitutedquinoline. In certain embodiments, R^(2C) is substituted quinoline. Incertain embodiments, R^(2C) is optionally substituted pyridine. Incertain embodiments, R^(2C) is pyridine substituted with a heterocyclicring. In some embodiments, R^(2C) is optionally substituted aliphatic.In certain embodiments, R^(2C) is unsubstituted aliphatic. In certainembodiments, R^(2C) is —CH₂-aryl. In certain embodiments, R^(2C) isbenzyl. In certain embodiments, R^(2C) is —CH₂-heteroaryl. In certainembodiments, R^(2C) is —CH₂-pyridyl. In some embodiments, R^(2C) is—C(═O)R^(A). In certain embodiments, R^(2C) is —C(═O)R^(A), whereinR^(A) is optionally substituted aliphatic. In certain embodiments, R² isacetyl. In certain embodiments, R^(2C) is —SO₂R^(A). In certainembodiments, R^(2C) is —SO₂R^(A), wherein R^(A) is optionallysubstituted aliphatic. In certain embodiments, R^(2C) is —SO₂CH₃.

As defined generally above, R^(3C) is selected from the group consistingof hydrogen, halo, optionally substituted aliphatic, optionallysubstituted carbocyclyl, optionally substituted aryl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂. In certainembodiments, R^(3C) is selected from the group consisting of hydrogen,halo, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted aryl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—OC(O)R^(A), —NR^(B)C(═O)R^(A), —NR^(B)C(═O)N(R^(B))₂, —SC(═O)R^(A),—C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(═O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂.

In certain embodiments, R^(3C) is hydrogen. In some embodiments, R^(3C)is not hydrogen. In some embodiments, R^(3C) is halo. In certainembodiments, R^(3C) is fluoro. In some embodiments, R^(3C) is optionallysubstituted aliphatic. In certain embodiments, R^(3C) is optionallysubstituted C₁₋₆ aliphatic. In certain embodiments, R^(3C) is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, R^(3C) is substitutedC₁₋₆ alkyl. In certain embodiments, R^(3C) is —CF₃, —CHF₂, or —CH₂F. Incertain embodiments, R^(3C) is unsubstituted C₁₋₆ alkyl. In certainembodiments, R^(3C) is methyl, ethyl, or propyl. In some embodiments,R^(3C) is —CN or —NO₂. In some embodiments, R^(3C) is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R^(3C) is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In some embodiments, R^(3C) isoptionally substituted aryl. In certain embodiments, R^(3C) isoptionally substituted phenyl. In certain embodiments, R^(3C) isunsubstituted phenyl. In certain embodiments, R^(3C) is halophenyl. Incertain embodiments, R^(3C) is fluorophenyl. In certain embodiments,R^(3C) is chlorophenyl. In some embodiments, R^(3C) is phenylsubstituted with optionally substituted C₁₋₆ alkyl. In some embodiments,R^(3C) is phenyl substituted with optionally substituted C₁₋₃ alkyl. Incertain embodiments, R^(3C) is phenyl substituted with methyl. Incertain embodiments, R^(3C) is phenyl substituted with —CH₂OH. In someembodiments, R^(3C) is phenyl substituted with a heterocyclic ring. Incertain embodiments, R^(3C) is phenyl substituted with morpholinyl. Incertain embodiments, R^(3C) is phenyl substituted withtetrahydropyranyl. In some embodiments, R^(3C) is optionally substitutedheteroaryl. In certain embodiments, R^(3C) is optionally substitutedquinoline. In certain embodiments, R^(3C) is unsubstituted quinoline. Incertain embodiments, R^(3C) is substituted quinoline. In certainembodiments, R^(3C) is optionally substituted pyridine. In certainembodiments, R^(3C) is pyridine substituted with a heterocyclic ring. Insome embodiments, R^(3C) is optionally substituted aliphatic. In certainembodiments, R^(3C) is unsubstituted aliphatic. In certain embodiments,R^(3C) is —CH₂-aryl. In certain embodiments, R^(3C) is benzyl. Incertain embodiments, R^(3C) is —CH₂-heteroaryl. In certain embodiments,R^(3C) is —CH₂-pyridyl.

As defined generally above, R^(4C) is selected from the group consistingof hydrogen, halo, or optionally substituted aliphatic. In certainembodiments, R^(4C) is hydrogen. In some embodiments, R^(4C) is nothydrogen. In some embodiments, R^(4C) is halo. In certain embodiments,R^(4C) is fluoro. In some embodiments, R^(4C) is optionally substitutedaliphatic. In certain embodiments, R^(4C) is optionally substituted C₁₋₆aliphatic. In certain embodiments, R^(4C) is optionally substituted C₁₋₆alkyl. In certain embodiments, R^(4C) is substituted C₁₋₆ alkyl. Incertain embodiments, R^(4C) is unsubstituted C₁₋₆ alkyl. In certainembodiments, R^(4C) is methyl, ethyl, or propyl.

As defined generally above, p is 0, 1, or 2. In certain embodiments, pis 0. In certain embodiments, p is 1. In certain embodiments, p is 2.

As defined generally above, L_(D) is —O—, —N(R)—, —C(R^(2A))(R^(3A))—,—O—CR^(2A)R^(3A), —N(R)—CR^(2A)R^(3A)—, —O—CR^(2A)R^(3A)—O—,—N(R)—CR^(2A)R^(3A)—O, —N(R)—CR^(2A)R^(3A)—N(R)—,—O—CR^(2A)R^(3A)—N(R)—, —CR^(2A)R^(3A)—O, —CR^(2A)R^(3A)—N(R),—O—CR^(2A)R^(3A)—CR⁹R¹⁰—, —N(R)—CR^(2A)R^(3A)—CR⁹R¹⁰—, —CR^(2A)R^(3A)CR⁹R¹⁰—O—, —CR^(2A)R^(3A)—CR⁹R—N(R)—, or —CR^(2A)R^(3A)—CR⁹R¹⁰—. Incertain embodiments, L_(D) is —O—, —N(R)—, or —CR^(2A)R^(3A)—, whereinR¹, R^(2A), and R^(3A) are as described herein. In certain embodiments,L_(D) is —O—. In some embodiments, L_(D) is —N(R)—. In certainembodiments, L_(D) is —NH—. In certain embodiments, L_(D) is —N(R)—,wherein R is optionally substituted C₁₋₆ aliphatic. In certainembodiments, L_(D) is —N(R)—, wherein R is optionally substituted C₁₋₆alkyl. In certain embodiments, L_(D) is —N(R)—, wherein R isunsubstituted C₁₋₆ alkyl. In certain embodiments, L_(D) is —N(R)—,wherein R is acetyl. In certain embodiments, L_(D) is —CH₂—O—. Incertain embodiments, L_(D) is —CR^(2A)R^(3A)—O—. In certain embodiments,L_(D) is —CR^(2A)R^(3A)—N(R)—. In certain embodiments, L_(D) is—CH₂—NH—.

As defined generally above, Ring A is a monocyclic or bicyclic,saturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur. Incertain embodiments, Ring A is aromatic. In certain embodiments, Ring Ais saturated. In certain embodiments, Ring A is monocyclic. In certainembodiments, Ring A is bicyclic.

In certain embodiments, Ring A is phenyl. In certain embodiments, Ring Ais a monocyclic heteroaryl having 1-3 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur. In certain embodiments, Ring A is a5- to 6-membered heteroaryl having 1-3 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In certain embodiments, RingA is a 5-membered heteroaryl having 1-3 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur (e.g., furanyl, thienyl,pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyrazolyl,isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl. In certainembodiments, Ring A is a 6-membered heteroaryl having 1-3 nitrogens(e.g., pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, triazinyl). Incertain embodiments, Ring A is pyridyl. In certain embodiments, Ring Ais pyrimidyl. In certain embodiments, Ring A is pyridazinyl. In someembodiments, Ring A is a carbocyclic ring. In some embodiments, Ring Ais a 3- to 8-membered saturated carbocyclic ring. In some embodiments,Ring A is a 3- to 8-membered heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur.

In certain embodiments, Ring A is a bicyclic saturated, partiallyunsaturated, or aromatic ring having 0-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In certain embodiments, RingA is an 8- to 12-membered bicyclic saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, Ring A is an 8- to10-membered bicyclic heteroaryl having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In certain embodiments, RingA is a 9-membered bicyclic heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur (e.g., indolyl,isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl,isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl,benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl,benzisothiazolyl, benzthiadiazolyl, indolizinyl). In certainembodiments, Ring A is a 10-membered bicyclic heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur(e.g., naphthyridinyl, quinolinyl, isoquinolinyl, quinoxalinyl,quinazolinyl. In certain embodiments, Ring A is selected from the groupconsisting of quinoline, benzimidazole, benzopyrazole, quinoxaline,tetrahydroquinoline, tetrahydroisoquinoline, naphthalene,tetrahydronaphthalene, 2,3-dihydrobenzo[b][1,4]dioxine, isoindole,2H-benzo[b][1,4]oxazin-3(4H)-one, 3,4-dihydro-2H-benzo[b][1,4]oxazine,and quinoxalin-2(1H)-one.

In some embodiments, q is 0. In some embodiments, q is 1.

As defined generally above, L₁ is a bond, —O—, —S—, —N(R)—, —C(O)—,—C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)—, —N(R)C(O)O—, —OC(O)N(R)—, —SO₂—,—SO₂N(R)—, —N(R)SO₂—, —OC(O)—, —C(O)O—, or an optionally substituted,straight or branched, C₁₋₆ aliphatic chain wherein one, two, or threemethylene units of L₁ are optionally and independently replaced by —O—,—S—, —N(R)—, —C(O)—, —C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)—,—N(R)C(O)O—, —OC(O)N(R)—, —SO₂—, —SO₂N(R)—, —N(R)SO₂—, —OC(O)—, or—C(O)O—. In some embodiments, L₁ is a bond. In some embodiments, L₁ is—O—, —S—, or —N(R)—. In some embodiments, L₁ is —C(O)—, —C(O)N(R)—, or—N(R)C(O)—. In some embodiments, L₁ is a C₁₋₆ aliphatic chain whereinone, two, or three methylene units of L₁ are optionally andindependently replaced by —O—, —S—, —N(R)—, —C(O)—, —C(O)N(R)—,—N(R)C(O)N(R)—, —N(R)C(O)—, —N(R)C(O)O—, —OC(O)N(R)—, —SO₂—, —SO₂N(R)—,—N(R)SO₂—, —OC(O)—, or —C(O)O—. In some embodiments, L₁ is a C₁₋₃aliphatic chain wherein one methylene unit of L₁ is optionally replacedby —O—, —S—, —N(R)—, —C(O)—, —C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)—,—N(R)C(O)O—, —OC(O)N(R)—, —SO₂—, —SO₂N(R)—, —N(R)SO₂—, —OC(O)—, or—C(O)O—. In some embodiments, L₁ is —CHNH—.

As defined generally above, Cy^(D) is an optionally substituted,monocyclic, bicyclic or tricyclic, saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, Cy^(D) isaromatic. In certain embodiments, Cy^(D) is saturated. In certainembodiments, Cy^(D) is monocyclic. In certain embodiments, Cy^(D) isbicyclic. In certain embodiments, Cy^(D) is tricyclic.

In certain embodiments, Cy^(D) is optionally substituted phenyl. Incertain embodiments, Cy^(D) is an optionally substituted 5- to6-membered heteroaryl having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, Cy^(D) is anoptionally substituted 5-membered heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur (e.g., furanyl,thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl,pyrazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl. Incertain embodiments, Cy^(D) is an optionally substituted 6-memberedheteroaryl having 1-3 nitrogens (e.g., pyridyl, pyrimidyl, pyridazinyl,pyrazinyl, triazinyl). In certain embodiments, Cy^(D) is optionallysubstituted pyrazole, optionally substituted pyridyl, or optionallysubstituted pyrimidyl. In some embodiments, Cy^(D) is an optionallysubstituted carbocyclic ring. In some embodiments, Cy^(D) is anoptionally substituted 3- to 8-membered saturated carbocyclic ring. Insome embodiments, Cy^(D) is an optionally substituted 3- to 8-memberedheterocyclic ring having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur.

In certain embodiments, Cy^(D) is an optionally substituted bicyclicsaturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur. Incertain embodiments, Cy^(D) is an optionally substituted 8- to12-membered bicyclic saturated, partially unsaturated, or aromatic ringhaving 0-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, Cy^(D) is an optionally substituted 8-to 10-membered bicyclic heteroaryl having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In certain embodiments,Cy^(D) is an optionally substituted 9- to 10-membered bicyclicheteroaryl having 1-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In certain embodiments, Cy^(D) is an optionallysubstituted 9-membered bicyclic heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur (e.g., indolyl,isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl,isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl,benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl,benzisothiazolyl, benzthiadiazolyl, indolizinyl). In certainembodiments, Cy^(D) is an optionally substituted 10-membered bicyclicheteroaryl having 1-3 heteroatoms independently selected from nitrogen,oxygen, and sulfur (e.g., naphthyridinyl, quinolinyl, isoquinolinyl,quinoxalinyl, quinazolinyl. In certain embodiments, Cy^(D) is optionallysubstituted indazole, optionally substituted quinoline, optionallysubstituted benzimidazole, optionally substituted benzothiazole,optionally substituted deazapurine, optionally substituted indole,optionally substituted purine, optionally substituted pyrazolopyridine,optionally substituted pyrrolopyridine, optionally substitutedpyrroloprimidine, optionally substituted imidazopyridine, or optionallysubstituted imidazopyridine.

As defined generally above, R⁹ and R¹⁰ are independently selected fromthe group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁹ and R¹⁰ are taken togetherwith their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring. In certain embodiments, R⁹ and R¹⁰ areindependently selected from the group consisting of hydrogen, halo, —CN,—NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted phenyl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—OC(O)R^(A), NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁹ and R¹⁰ are taken togetherwith their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring.

In certain embodiments, R⁹ is hydrogen. In some embodiments, R⁹ is nothydrogen. In some embodiments, R⁹ is halo. In certain embodiments, R⁹ isfluoro. In some embodiments, R⁹ is optionally substituted aliphatic. Incertain embodiments, R⁹ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R⁹ is optionally substituted C₁₋₆ alkyl. In certainembodiments, R⁹ is substituted C₁₋₆ alkyl. In certain embodiments, R⁹ is—CF₃, CHF₂, or CH₂F. In certain embodiments, R⁹ is unsubstituted C₁₋₆alkyl. In certain embodiments, R⁹ is methyl, ethyl, or propyl. In someembodiments, R⁹ is —CN or —NO₂. In some embodiments, R⁹ is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R⁹ is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R⁹ is—N(R^(B))₂. In certain embodiments, R⁹ is —NHR^(B). In certainembodiments, R⁹ is —NH₂. In certain embodiments, R⁹ is —OR^(A). Incertain embodiments, R⁹ is —OH.

In certain embodiments, R¹⁰ is hydrogen. In some embodiments, R¹⁰ is nothydrogen. In some embodiments, R¹⁰ is halo. In certain embodiments, R¹⁰is fluoro. In some embodiments, R¹⁰ is optionally substituted aliphatic.In certain embodiments, R¹⁰ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R¹⁰ is optionally substituted C₁₋₆ alkyl. Incertain embodiments, R¹⁰ is substituted C₁₋₆ alkyl. In certainembodiments, R¹⁰ is —CF₃, CHF₂, or CH₂F. In certain embodiments, R¹⁰ isunsubstituted C₁₋₆ alkyl. In certain embodiments, R¹⁰ is methyl, ethyl,or propyl. In some embodiments, R¹⁰ is —CN or —NO₂. In some embodiments,R¹⁰ is optionally substituted carbocyclyl, optionally substitutedphenyl, optionally substituted heterocyclyl, or optionally substitutedheteroaryl. In some embodiments, R¹⁰ is —OR^(A), —N(R^(B))₂, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A),—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R¹⁰ is—N(R^(B))₂. In certain embodiments, R¹⁰ is —NHR^(B). In certainembodiments, R¹⁰ is —NH₂. In certain embodiments, R¹⁰ is —OR^(A). Incertain embodiments, R¹⁰ is —OH.

In some embodiments, R⁹ and R¹⁰ are the same. In some embodiments, R⁹and R¹⁰ are different. In some embodiments, R⁹ and R¹⁰ are eachhydrogen. In some embodiments, R⁹ is hydrogen and R¹⁰ is not hydrogen.In some embodiments, R⁹ is hydrogen and R¹⁰ is optionally substitutedaliphatic. In some embodiments, R⁹ is hydrogen and R¹⁰ is C₁₋₆ alkyl. Insome embodiments, R⁹ is hydrogen and R¹⁰ is methyl. In some embodiments,R⁹ is hydrogen and R¹⁰ is ethyl or propyl. In certain embodiments, R⁹and hydrogen and R¹⁰ is —CF₃, CHF₂, or CH₂F. In some embodiments, R⁹ ishydrogen and R¹⁰ is —N(R^(B))₂ or —OR^(A). In some embodiments, R⁹ ishydrogen and R¹⁰ is —NH₂. In some embodiments, R⁹ is hydrogen and R¹⁰ is—OH. In some embodiments, R⁹ and R¹⁰ are not hydrogen. In someembodiments, R⁹ and R¹⁰ are independently optionally substitutedaliphatic. In some embodiments, R⁹ and R¹⁰ are methyl. In someembodiments, R⁹ and R¹⁰ are taken together with their intervening atomsto form an optionally substituted carbocyclic or heterocyclic ring.

As defined generally above, each R^(x) is independently selected fromthe group consisting of halo, —CN, optionally substituted aliphatic, and—OR′. In certain embodiments, at least one R^(x) is halo. In certainembodiments, at least one R^(x) is fluoro. In certain embodiments, atleast one R^(x) is —CN. In certain embodiments, at least one R^(x) isoptionally substituted aliphatic. In certain embodiments, at least oneR^(x) is optionally substituted C₁₋₆ alkyl. In certain embodiments, atleast one R^(x) is methyl. In certain embodiments, at least one R^(x) is—CF₃. In certain embodiments, at least one R^(x) is —OR′. In certainembodiments, R^(x) is not —OR′. In certain embodiments, at least oneR^(x) is —OCH₃. In certain embodiments, R^(x) is not —OCH₃.

One of ordinary skill in the art will appreciate that an R^(x) group canbe attached anywhere on the isoindoline ring. In certain embodiments, anR^(x) group is attached to the benzene portion of the isoindoline ring.In certain embodiments, an R^(x) group is attached to the dihydropyrroleportion of the isoindoline ring. In certain embodiments, R^(x) groupsare attached to both the benzene portion and the dihydropyrrole portionof the isoindoline ring. See, for example, the structures shown below:

As defined generally above, n is 0, 1, 2, 3, 4, 5, 6, 7, or 8. Incertain embodiments, n is 0. In certain embodiments, n is 1. In certainembodiments, n is 2.

As defined generally above, k is 0, 1, 2, 3, or 4. In some embodiments,k is 0. In some embodiments, k is 1. In some embodiments, k is 2.

In certain embodiments, a provided compound is a compound listed inTable 1, or a pharmaceutically acceptable salt thereof.

TABLE 1 Exemplary Compounds Cmpd LC-MS m/z No Structure Exact mass (M +H) 1

391.1896 392.1 2

311.1634 312.2 3

391.1896 392.1 4

377.1739 378.1 5

391.1896 392.1 6

391.1896 392.1 7

392.1848 393.2 8

395.2209 396.2 9

399.1947 400.2 10

397.2002 398.2 11

411.2158 412.2 12

404.1406 405.2 13

373.179 374.2 14

399.1947 400.2 15

433.1671 434.1 16

377.1739 378.1 17

399.1947 400.2 18

505.2365 506.2 19

377.1739 378.1 20

366.2307 367.2 21

433.1671 434.2 22

397.2002 398.1 23

433.1671 24

395.2209 396.2 25

380.1848 26

397.2002 398.2 27

380.1848 381.1 28

395.2209 396.2 29

397.2114 398.1 30

396.2161 31

397.2002 398.2 32

381.2165 382.1 33

459.194 460.3 34

396.2161

In certain embodiments, a provided compound inhibits PRMT5. In certainembodiments, a provided compound inhibits wild-type PRMT5. In certainembodiments, a provided compound inhibits a mutant PRMT5. In certainembodiments, a provided compound inhibits PRMT5, e.g., as measured in anassay described herein. In certain embodiments, the PRMT5 is from ahuman. In certain embodiments, a provided compound inhibits PRMT5 at anIC₅₀ less than or equal to 10 μM. In certain embodiments, a providedcompound inhibits PRMT5 at an IC₅₀ less than or equal to 1 μM. Incertain embodiments, a provided compound inhibits PRMT5 at an IC₅₀ lessthan or equal to 0.1 μM. In certain embodiments, a provided compoundinhibits PRMT5 in a cell at an EC₅₀ less than or equal to 10 μM. Incertain embodiments, a provided compound inhibits PRMT5 in a cell at anEC₅₀ less than or equal to 1 μM. In certain embodiments, a providedcompound inhibits PRMT5 in a cell at an EC₅₀ less than or equal to 0.1μM. In certain embodiments, a provided compound inhibits cellproliferation at an EC₅₀ less than or equal to 10 μM. In certainembodiments, a provided compound inhibits cell proliferation at an EC₅₀less than or equal to 1 μM. In certain embodiments, a provided compoundinhibits cell proliferation at an EC₅₀ less than or equal to 0.1 μM. Insome embodiments, a provided compound is selective for PRMT5 over othermethyltransferases. In certain embodiments, a provided compound is atleast about 10-fold selective, at least about 20-fold selective, atleast about 30-fold selective, at least about 40-fold selective, atleast about 50-fold selective, at least about 60-fold selective, atleast about 70-fold selective, at least about 80-fold selective, atleast about 90-fold selective, or at least about 100-fold selective forPRMT5 relative to one or more other methyltransferases.

It will be understood by one of ordinary skill in the art that the PRMT5can be wild-type PRMT5, or any mutant or variant of PRMT5.

In certain embodiments, the PRMT5 is isoform A (GenBank accession no.NP006100) (SEQ ID NO.:1):

MAAMAVGGAG GSRVSSGRDL NCVPEIADTL GAVAKQGFDFLCMPVFHPRF KREFIQEPAK NRPGPQTRSD LLLSGRDWNTLIVGKLSPWI RPDSKVEKIR RNSEAAMLQE LNFGAYLGLPAFLLPLNQED NTNLARVLTN HIHTGHHSSM FWMRVPLVAPEDLRDDIIEN APTTHTEEYS GEEKTWMWWH NFRTLCDYSKRIAVALEIGA DLPSNHVIDR WLGEPIKAAI LPTSIFLTNKKGFPVLSKMH QRLIFRLLKL EVQFIITGTN HHSEKEFCSYLQYLEYLSQN RPPPNAYELF AKGYEDYLQS PLQPLMDNLESQTYEVFEKD PIKYSQYQQA IYKCLLDRVP EEEKDTNVQVLMVLGAGRGP LVNASLRAAK QADRRIKLYA VEKNPNAVVTLENWQFEEWG SQVTVVSSDM REWVAPEKAD IIVSELLGSFADNELSPECL DGAQHFLKDD GVSIPGEYTS FLAPISSSKLYNEVRACREK DRDPEAQFEM PYVVRLHNFH QLSAPQPCFTFSHPNRDPMI DNNRYCTLEF PVEVNTVLHG FAGYFETVLYQDITLSIRPE THSPGMFSWF PILFPIKQPI TVREGQTICVRFWRCSNSKK VWYEWAVTAP VCSAIHNPTG RSYTIGL

In certain embodiments, the PRMT5 is isoform B (GenBank accession no.NP001034708) (SEQ ID NO.:2)

MRGPNSGTEK GRLVIPEKQG FDFLCMPVFH PRFKREFIQEPAKNRPGPQT RSDLLLSGRD WNTLIVGKLS PWIRPDSKVEKIRRNSEAAM LQELNFGAYL GLPAFLLPLN QEDNTNLARVLTNHIHTGHH SSMFWMRVPL VAPEDLRDDI IENAPTTHTEEYSGEEKTWM WWHNFRTLCD YSKRIAVALE IGADLPSNHVIDRWLGEPIK AAILPTSIFL TNKKGFPVLS KMHQRLIFRLLKLEVQFIIT GTNHHSEKEF CSYLQYLEYL SQNRPPPNAYELFAKGYEDY LQSPLQPLMD NLESQTYEVF EKDPIKYSQYQQAIYKCLLD RVPEEEKDTN VQVLMVLGAG RGPLVNASLRAAKQADRRIK LYAVEKNPNA VVTLENWQFE EWGSQVTVVSSDMREWVAPE KADIIVSELL GSFADNELSP ECLDGAQHFLKDDGVSIPGE YTSFLAPISS SKLYNEVRAC REKDRDPEAQFEMPYVVRLH NFHQLSAPQP CFTFSHPNRD PMIDNNRYCTLEFPVEVNTV LHGFAGYFET VLYQDITLSI RPETHSPGMFSWFPILFPIK QPITVREGQT ICVRFWRCSN SKKVWYEWAV TAPVCSAIHN PTGRSYTIGL

In certain embodiments, the PRMT5 is transcript variant 1 (GenBankaccession no. NM_(—)006109).

The present disclosure provides pharmaceutical compositions comprising acompound described herein, e.g., a compound of Formula (I), or apharmaceutically acceptable salt thereof, as described herein, andoptionally a pharmaceutically acceptable excipient. It will beunderstood by one of ordinary skill in the art that the compoundsdescribed herein, or salts thereof, may be present in various forms,such as hydrates, solvates, or polymorphs. In certain embodiments, aprovided composition comprises two or more compounds described herein.In certain embodiments, a compound described herein, or apharmaceutically acceptable salt thereof, is provided in an effectiveamount in the pharmaceutical composition. In certain embodiments, theeffective amount is a therapeutically effective amount. In certainembodiments, the effective amount is an amount effective for inhibitingPRMT5. In certain embodiments, the effective amount is an amounteffective for treating a PRMT5-mediated disorder. In certainembodiments, the effective amount is a prophylactically effectiveamount. In certain embodiments, the effective amount is an amounteffective to prevent a PRMT5-mediated disorder.

Pharmaceutically acceptable excipients include any and all solvents,diluents, or other liquid vehicles, dispersions, suspension aids,surface active agents, isotonic agents, thickening or emulsifyingagents, preservatives, solid binders, lubricants, and the like, assuited to the particular dosage form desired. General considerations informulation and/or manufacture of pharmaceutical compositions agents canbe found, for example, in Remington's Pharmaceutical Sciences, SixteenthEdition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980), andRemington: The Science and Practice of Pharmacy, 21st Edition(Lippincott Williams & Wilkins, 2005).

Pharmaceutical compositions described herein can be prepared by anymethod known in the art of pharmacology. In general, such preparatorymethods include the steps of bringing a compound described herein (the“active ingredient”) into association with a carrier and/or one or moreother accessory ingredients, and then, if necessary and/or desirable,shaping and/or packaging the product into a desired single- ormulti-dose unit.

Pharmaceutical compositions can be prepared, packaged, and/or sold inbulk, as a single unit dose, and/or as a plurality of single unit doses.As used herein, a “unit dose” is discrete amount of the pharmaceuticalcomposition comprising a predetermined amount of the active ingredient.The amount of the active ingredient is generally equal to the dosage ofthe active ingredient which would be administered to a subject and/or aconvenient fraction of such a dosage such as, for example, one-half orone-third of such a dosage.

Relative amounts of the active ingredient, the pharmaceuticallyacceptable excipient, and/or any additional ingredients in apharmaceutical composition of the present disclosure will vary,depending upon the identity, size, and/or condition of the subjecttreated and further depending upon the route by which the composition isto be administered. By way of example, the composition may comprisebetween 0.1% and 100% (w/w) active ingredient.

Pharmaceutically acceptable excipients used in the manufacture ofprovided pharmaceutical compositions include inert diluents, dispersingand/or granulating agents, surface active agents and/or emulsifiers,disintegrating agents, binding agents, preservatives, buffering agents,lubricating agents, and/or oils. Excipients such as cocoa butter andsuppository waxes, coloring agents, coating agents, sweetening,flavoring, and perfuming agents may also be present in the composition.

Exemplary diluents include calcium carbonate, sodium carbonate, calciumphosphate, dicalcium phosphate, calcium sulfate, calcium hydrogenphosphate, sodium phosphate lactose, sucrose, cellulose,microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodiumchloride, dry starch, cornstarch, powdered sugar, and mixtures thereof.

Exemplary granulating and/or dispersing agents include potato starch,corn starch, tapioca starch, sodium starch glycolate, clays, alginicacid, guar gum, citrus pulp, agar, bentonite, cellulose and woodproducts, natural sponge, cation-exchange resins, calcium carbonate,silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone)(crospovidone), sodium carboxymethyl starch (sodium starch glycolate),carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose(croscarmellose), methylcellulose, pregelatinized starch (starch 1500),microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate,quaternary ammonium compounds, and mixtures thereof.

Exemplary surface active agents and/or emulsifiers include naturalemulsifiers (e.g., acacia, agar, alginic acid, sodium alginate,tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk,casein, wool fat, cholesterol, wax, and lecithin), colloidal clays(e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminumsilicate)), long chain amino acid derivatives, high molecular weightalcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetinmonostearate, ethylene glycol distearate, glyceryl monostearate, andpropylene glycol monostearate, polyvinyl alcohol), carbomers (e.g.,carboxy polymethylene, polyacrylic acid, acrylic acid polymer, andcarboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g.,carboxymethylcellulose sodium, powdered cellulose, hydroxymethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylenesorbitan monolaurate (Tween 20), polyoxyethylene sorbitan (Tween 60),polyoxyethylene sorbitan monooleate (Tween 80), sorbitan monopalmitate(Span 40), sorbitan monostearate (Span 60], sorbitan tristearate (Span65), glyceryl monooleate, sorbitan monooleate (Span 80)),polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj 45),polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil,polyoxymethylene stearate, and Solutol), sucrose fatty acid esters,polyethylene glycol fatty acid esters (e.g., Cremophor™),polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether (Brij 30)),poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamineoleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyllaurate, sodium lauryl sulfate, Pluronic F68, Poloxamer 188, cetrimoniumbromide, cetylpyridinium chloride, benzalkonium chloride, docusatesodium, and/or mixtures thereof.

Exemplary binding agents include starch (e.g., cornstarch and starchpaste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin,molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums(e.g., acacia, sodium alginate, extract of Irish moss, panwar gum,ghatti gum, mucilage of isapol husks, carboxymethylcellulose,methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose,cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate(Veegum), and larch arabogalactan), alginates, polyethylene oxide,polyethylene glycol, inorganic calcium salts, silicic acid,polymethacrylates, waxes, water, alcohol, and/or mixtures thereof.

Exemplary preservatives include antioxidants, chelating agents,antimicrobial preservatives, antifungal preservatives, alcoholpreservatives, acidic preservatives, and other preservatives.

Exemplary antioxidants include alpha tocopherol, ascorbic acid, acorbylpalmitate, butylated hydroxyanisole, butylated hydroxytoluene,monothioglycerol, potassium metabisulfite, propionic acid, propylgallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, andsodium sulfite.

Exemplary chelating agents include ethylenediaminetetraacetic acid(EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodiumedetate, trisodium edetate, calcium disodium edetate, dipotassiumedetate, and the like), citric acid and salts and hydrates thereof(e.g., citric acid monohydrate), fumaric acid and salts and hydratesthereof, malic acid and salts and hydrates thereof, phosphoric acid andsalts and hydrates thereof, and tartaric acid and salts and hydratesthereof. Exemplary antimicrobial preservatives include benzalkoniumchloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide,cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol,chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea,phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate,propylene glycol, and thimerosal.

Exemplary antifungal preservatives include butyl paraben, methylparaben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoicacid, potassium benzoate, potassium sorbate, sodium benzoate, sodiumpropionate, and sorbic acid.

Exemplary alcohol preservatives include ethanol, polyethylene glycol,phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate,and phenylethyl alcohol.

Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E,beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbicacid, sorbic acid, and phytic acid.

Other preservatives include tocopherol, tocopherol acetate, deteroximemesylate, cetrimide, butylated hydroxyanisol (BHA), butylatedhydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS),sodium lauryl ether sulfate (SLES), sodium bisulfite, sodiummetabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus,Phenonip, methylparaben, Germall 115, Germaben II, Neolone, Kathon, andEuxyl. In certain embodiments, the preservative is an anti-oxidant. Inother embodiments, the preservative is a chelating agent.

Exemplary buffering agents include citrate buffer solutions, acetatebuffer solutions, phosphate buffer solutions, ammonium chloride, calciumcarbonate, calcium chloride, calcium citrate, calcium glubionate,calcium gluceptate, calcium gluconate, D-gluconic acid, calciumglycerophosphate, calcium lactate, propanoic acid, calcium levulinate,pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasiccalcium phosphate, calcium hydroxide phosphate, potassium acetate,potassium chloride, potassium gluconate, potassium mixtures, dibasicpotassium phosphate, monobasic potassium phosphate, potassium phosphatemixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodiumcitrate, sodium lactate, dibasic sodium phosphate, monobasic sodiumphosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide,aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline,Ringer's solution, ethyl alcohol, and mixtures thereof.

Exemplary lubricating agents include magnesium stearate, calciumstearate, stearic acid, silica, talc, malt, glyceryl behanate,hydrogenated vegetable oils, polyethylene glycol, sodium benzoate,sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate,sodium lauryl sulfate, and mixtures thereof.

Exemplary natural oils include almond, apricot kernel, avocado, babassu,bergamot, black current seed, borage, cade, camomile, canola, caraway,carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee,corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed,geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate,jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba, macademianut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange,orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed,pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood,sasquana, savoury, sea buckthorn, sesame, shea butter, silicone,soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, andwheat germ oils. Exemplary synthetic oils include, but are not limitedto, butyl stearate, caprylic triglyceride, capric triglyceride,cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate,mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and mixturesthereof.

Liquid dosage forms for oral and parenteral administration includepharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active ingredients,the liquid dosage forms may comprise inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed,groundnut, corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can include adjuvants such as wetting agents, emulsifyingand suspending agents, sweetening, flavoring, and perfuming agents. Incertain embodiments for parenteral administration, the compoundsdescribed herein are mixed with solubilizing agents such as Cremophor™,alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins,polymers, and mixtures thereof.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions can be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation can be a sterile injectable solution,suspension or emulsion in a nontoxic parenterally acceptable diluent orsolvent, for example, as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that can be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This can be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Compositions for rectal or vaginal administration are typicallysuppositories which can be prepared by mixing the compounds describedherein with suitable non-irritating excipients or carriers such as cocoabutter, polyethylene glycol or a suppository wax which are solid atambient temperature but liquid at body temperature and therefore melt inthe rectum or vaginal cavity and release the active ingredient.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activeingredient is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may comprise buffering agents.

Solid compositions of a similar type can be employed as fillers in softand hard-filled gelatin capsules using such excipients as lactose ormilk sugar as well as high molecular weight polyethylene glycols and thelike. The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes. Solid compositions of asimilar type can be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polyethylene glycols and the like.

The active ingredient can be in micro-encapsulated form with one or moreexcipients as noted above. The solid dosage forms of tablets, dragees,capsules, pills, and granules can be prepared with coatings and shellssuch as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active ingredient can be admixed with at least oneinert diluent such as sucrose, lactose, or starch. Such dosage forms maycomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets, and pills, the dosage forms may comprise bufferingagents. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes.

Dosage forms for topical and/or transdermal administration of a providedcompound may include ointments, pastes, creams, lotions, gels, powders,solutions, sprays, inhalants and/or patches. Generally, the activeingredient is admixed under sterile conditions with a pharmaceuticallyacceptable carrier and/or any desired preservatives and/or buffers ascan be required. Additionally, the present disclosure encompasses theuse of transdermal patches, which often have the added advantage ofproviding controlled delivery of an active ingredient to the body. Suchdosage forms can be prepared, for example, by dissolving and/ordispensing the active ingredient in the proper medium. Alternatively oradditionally, the rate can be controlled by either providing a ratecontrolling membrane and/or by dispersing the active ingredient in apolymer matrix and/or gel.

Suitable devices for use in delivering intradermal pharmaceuticalcompositions described herein include short needle devices such as thosedescribed in U.S. Pat. Nos. 4,886,499; 5,190,521; 5,328,483; 5,527,288;4,270,537; 5,015,235; 5,141,496; and 5,417,662. Intradermal compositionscan be administered by devices which limit the effective penetrationlength of a needle into the skin, such as those described in PCTpublication WO 99/34850 and functional equivalents thereof. Jetinjection devices which deliver liquid vaccines to the dermis via aliquid jet injector and/or via a needle which pierces the stratumcorneum and produces a jet which reaches the dermis are suitable. Jetinjection devices are described, for example, in U.S. Pat. Nos.5,480,381; 5,599,302; 5,334,144; 5,993,412; 5,649,912; 5,569,189;5,704,911; 5,383,851; 5,893,397; 5,466,220; 5,339,163; 5,312,335;5,503,627; 5,064,413; 5,520,639; 4,596,556; 4,790,824; 4,941,880;4,940,460; and PCT publications WO 97/37705 and WO 97/13537. Ballisticpowder/particle delivery devices which use compressed gas to acceleratevaccine in powder form through the outer layers of the skin to thedermis are suitable. Alternatively or additionally, conventionalsyringes can be used in the classical mantoux method of intradermaladministration.

Formulations suitable for topical administration include, but are notlimited to, liquid and/or semi liquid preparations such as liniments,lotions, oil in water and/or water in oil emulsions such as creams,ointments and/or pastes, and/or solutions and/or suspensions.Topically-administrable formulations may, for example, comprise fromabout 1% to about 10% (w/w) active ingredient, although theconcentration of the active ingredient can be as high as the solubilitylimit of the active ingredient in the solvent. Formulations for topicaladministration may further comprise one or more of the additionalingredients described herein.

A provided pharmaceutical composition can be prepared, packaged, and/orsold in a formulation suitable for pulmonary administration via thebuccal cavity. Such a formulation may comprise dry particles whichcomprise the active ingredient and which have a diameter in the rangefrom about 0.5 to about 7 nanometers or from about 1 to about 6nanometers. Such compositions are conveniently in the form of drypowders for administration using a device comprising a dry powderreservoir to which a stream of propellant can be directed to dispersethe powder and/or using a self propelling solvent/powder dispensingcontainer such as a device comprising the active ingredient dissolvedand/or suspended in a low-boiling propellant in a sealed container. Suchpowders comprise particles wherein at least 98% of the particles byweight have a diameter greater than 0.5 nanometers and at least 95% ofthe particles by number have a diameter less than 7 nanometers.Alternatively, at least 95% of the particles by weight have a diametergreater than 1 nanometer and at least 90% of the particles by numberhave a diameter less than 6 nanometers. Dry powder compositions mayinclude a solid fine powder diluent such as sugar and are convenientlyprovided in a unit dose form.

Low boiling propellants generally include liquid propellants having aboiling point of below 65° F. at atmospheric pressure. Generally thepropellant may constitute 50 to 99.9% (w/w) of the composition, and theactive ingredient may constitute 0.1 to 20% (w/w) of the composition.The propellant may further comprise additional ingredients such as aliquid non-ionic and/or solid anionic surfactant and/or a solid diluent(which may have a particle size of the same order as particlescomprising the active ingredient).

Pharmaceutical compositions formulated for pulmonary delivery mayprovide the active ingredient in the form of droplets of a solutionand/or suspension. Such formulations can be prepared, packaged, and/orsold as aqueous and/or dilute alcoholic solutions and/or suspensions,optionally sterile, comprising the active ingredient, and mayconveniently be administered using any nebulization and/or atomizationdevice. Such formulations may further comprise one or more additionalingredients including, but not limited to, a flavoring agent such assaccharin sodium, a volatile oil, a buffering agent, a surface activeagent, and/or a preservative such as methylhydroxybenzoate. The dropletsprovided by this route of administration may have an average diameter inthe range from about 0.1 to about 200 nanometers.

Formulations described herein as being useful for pulmonary delivery areuseful for intranasal delivery of a pharmaceutical composition. Anotherformulation suitable for intranasal administration is a coarse powdercomprising the active ingredient and having an average particle fromabout 0.2 to 500 micrometers. Such a formulation is administered byrapid inhalation through the nasal passage from a container of thepowder held close to the nares.

Formulations for nasal administration may, for example, comprise fromabout as little as 0.1% (w/w) and as much as 100% (w/w) of the activeingredient, and may comprise one or more of the additional ingredientsdescribed herein. A provided pharmaceutical composition can be prepared,packaged, and/or sold in a formulation for buccal administration. Suchformulations may, for example, be in the form of tablets and/or lozengesmade using conventional methods, and may contain, for example, 0.1 to20% (w/w) active ingredient, the balance comprising an orallydissolvable and/or degradable composition and, optionally, one or moreof the additional ingredients described herein. Alternately,formulations for buccal administration may comprise a powder and/or anaerosolized and/or atomized solution and/or suspension comprising theactive ingredient. Such powdered, aerosolized, and/or aerosolizedformulations, when dispersed, may have an average particle and/ordroplet size in the range from about 0.1 to about 200 nanometers, andmay further comprise one or more of the additional ingredients describedherein.

A provided pharmaceutical composition can be prepared, packaged, and/orsold in a formulation for ophthalmic administration. Such formulationsmay, for example, be in the form of eye drops including, for example, a0.1/1.0% (w/w) solution and/or suspension of the active ingredient in anaqueous or oily liquid carrier. Such drops may further comprisebuffering agents, salts, and/or one or more other of the additionalingredients described herein. Other opthalmically-administrableformulations which are useful include those which comprise the activeingredient in microcrystalline form and/or in a liposomal preparation.Ear drops and/or eye drops are contemplated as being within the scope ofthis disclosure.

Although the descriptions of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions which aresuitable for administration to humans, it will be understood by theskilled artisan that such compositions are generally suitable foradministration to animals of all sorts. Modification of pharmaceuticalcompositions suitable for administration to humans in order to renderthe compositions suitable for administration to various animals is wellunderstood, and the ordinarily skilled veterinary pharmacologist candesign and/or perform such modification with ordinary experimentation.

Compounds provided herein are typically formulated in dosage unit formfor ease of administration and uniformity of dosage. It will beunderstood, however, that the total daily usage of provided compositionswill be decided by the attending physician within the scope of soundmedical judgment. The specific therapeutically effective dose level forany particular subject or organism will depend upon a variety of factorsincluding the disease, disorder, or condition being treated and theseverity of the disorder; the activity of the specific active ingredientemployed; the specific composition employed; the age, body weight,general health, sex and diet of the subject; the time of administration,route of administration, and rate of excretion of the specific activeingredient employed; the duration of the treatment; drugs used incombination or coincidental with the specific active ingredientemployed; and like factors well known in the medical arts.

The compounds and compositions provided herein can be administered byany route, including enteral (e.g., oral), parenteral, intravenous,intramuscular, intra-arterial, intramedullary, intrathecal,subcutaneous, intraventricular, transdermal, interdermal, rectal,intravaginal, intraperitoneal, topical (as by powders, ointments,creams, and/or drops), mucosal, nasal, bucal, sublingual; byintratracheal instillation, bronchial instillation, and/or inhalation;and/or as an oral spray, nasal spray, and/or aerosol. Specificallycontemplated routes are oral administration, intravenous administration(e.g., systemic intravenous injection), regional administration viablood and/or lymph supply, and/or direct administration to an affectedsite. In general the most appropriate route of administration willdepend upon a variety of factors including the nature of the agent(e.g., its stability in the environment of the gastrointestinal tract),and/or the condition of the subject (e.g., whether the subject is ableto tolerate oral administration).

The exact amount of a compound required to achieve an effective amountwill vary from subject to subject, depending, for example, on species,age, and general condition of a subject, severity of the side effects ordisorder, identity of the particular compound(s), mode ofadministration, and the like. The desired dosage can be delivered threetimes a day, two times a day, once a day, every other day, every thirdday, every week, every two weeks, every three weeks, or every fourweeks. In certain embodiments, the desired dosage can be delivered usingmultiple administrations (e.g., two, three, four, five, six, seven,eight, nine, ten, eleven, twelve, thirteen, fourteen, or moreadministrations).

In certain embodiments, an effective amount of a compound foradministration one or more times a day to a 70 kg adult human maycomprise about 0.0001 mg to about 3000 mg, about 0.0001 mg to about 2000mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg,about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about1000 mg, or about 100 mg to about 1000 mg, of a compound per unit dosageform.

In certain embodiments, a compound described herein may be administeredat dosage levels sufficient to deliver from about 0.001 mg/kg to about1000 mg/kg, from about 0.01 mg/kg to about mg/kg, from about 0.1 mg/kgto about 40 mg/kg, from about 0.5 mg/kg to about 30 mg/kg, from about0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, orfrom about 1 mg/kg to about 25 mg/kg, of subject body weight per day,one or more times a day, to obtain the desired therapeutic effect.

In some embodiments, a compound described herein is administered one ormore times per day, for multiple days. In some embodiments, the dosingregimen is continued for days, weeks, months, or years.

It will be appreciated that dose ranges as described herein provideguidance for the administration of provided pharmaceutical compositionsto an adult. The amount to be administered to, for example, a child oran adolescent can be determined by a medical practitioner or personskilled in the art and can be lower or the same as that administered toan adult.

It will be also appreciated that a compound or composition, as describedherein, can be administered in combination with one or more additionaltherapeutically active agents. In certain embodiments, a compound orcomposition provided herein is administered in combination with one ormore additional therapeutically active agents that improve itsbioavailability, reduce and/or modify its metabolism, inhibit itsexcretion, and/or modify its distribution within the body. It will alsobe appreciated that the therapy employed may achieve a desired effectfor the same disorder, and/or it may achieve different effects.

The compound or composition can be administered concurrently with, priorto, or subsequent to, one or more additional therapeutically activeagents. In certain embodiments, the additional therapeutically activeagent is a compound of Formula (I). In certain embodiments, theadditional therapeutically active agent is not a compound of Formula(I). In general, each agent will be administered at a dose and/or on atime schedule determined for that agent. In will further be appreciatedthat the additional therapeutically active agent utilized in thiscombination can be administered together in a single composition oradministered separately in different compositions. The particularcombination to employ in a regimen will take into account compatibilityof a provided compound with the additional therapeutically active agentand/or the desired therapeutic effect to be achieved. In general, it isexpected that additional therapeutically active agents utilized incombination be utilized at levels that do not exceed the levels at whichthey are utilized individually. In some embodiments, the levels utilizedin combination will be lower than those utilized individually.

Exemplary additional therapeutically active agents include, but are notlimited to, small organic molecules such as drug compounds (e.g.,compounds approved by the U.S. Food and Drug Administration as providedin the Code of Federal Regulations (CFR)), peptides, proteins,carbohydrates, monosaccharides, oligosaccharides, polysaccharides,nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides orproteins, small molecules linked to proteins, glycoproteins, steroids,nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides,antisense oligonucleotides, lipids, hormones, vitamins, and cells.

Also encompassed by the present discosure are kits (e.g., pharmaceuticalpacks). The kits provided may comprise a provided pharmaceuticalcomposition or compound and a container (e.g., a vial, ampule, bottle,syringe, and/or dispenser package, or other suitable container). In someembodiments, provided kits may optionally further include a secondcontainer comprising a pharmaceutical excipient for dilution orsuspension of a provided pharmaceutical composition or compound. In someembodiments, a provided pharmaceutical composition or compound providedin the container and the second container are combined to form one unitdosage form. In some embodiments, a provided kits further includesinstructions for use.

Compounds and compositions described herein are generally useful for theinhibition of PRMT5. In some embodiments, methods of treatingPRMT5-mediated disorder in a subject are provided which compriseadministering an effective amount of a compound described herein (e.g.,a compound of Formula (I)), or a pharmaceutically acceptable saltthereof), to a subject in need of treatment. In certain embodiments, theeffective amount is a therapeutically effective amount. In certainembodiments, the effective amount is a prophylactically effectiveamount. In certain embodiments, the subject is suffering from aPRMT5-mediated disorder. In certain embodiments, the subject issusceptible to a PRMT5-mediated disorder.

As used herein, the term “PRMT5-mediated disorder” means any disease,disorder, or other pathological condition in which PRMT5 is known toplay a role. Accordingly, in some embodiments, the present disclosurerelates to treating or lessening the severity of one or more diseases inwhich PRMT5 is known to play a role.

In some embodiments, the present disclosure provides a method ofinhibiting PRMT5 comprising contacting PRMT5 with an effective amount ofa compound described herein (e.g., a compound of Formula (I)), or apharmaceutically acceptable salt thereof. The PRMT5 may be purified orcrude, and may be present in a cell, tissue, or subject. Thus, suchmethods encompass both inhibition of in vitro and in vivo PRMT5activity. In certain embodiments, the method is an in vitro method,e.g., such as an assay method. It will be understood by one of ordinaryskill in the art that inhibition of PRMT5 does not necessarily requirethat all of the PRMT5 be occupied by an inhibitor at once. Exemplarylevels of inhibition of PRMT5 include at least 10% inhibition, about 10%to about 25% inhibition, about 25% to about 50% inhibition, about 50% toabout 75% inhibition, at least 50% inhibition, at least 75% inhibition,about 80% inhibition, about 90% inhibition, and greater than 90%inhibition.

In some embodiments, provided is a method of inhibiting PRMT5 activityin a subject in need thereof comprising administering to the subject aneffective amount of a compound described herein (e.g., a compound ofFormula (I)), or a pharmaceutically acceptable salt thereof, or apharmaceutical composition thereof.

In certain embodiments, provided is a method of altering gene expressionin a cell which comprises contacting a cell with an effective amount ofa compound of Formula (I), or a pharmaceutically acceptable saltthereof. In certain embodiments, the cell in culture in vitro. Incertain embodiments, the cell is in an animal, e.g., a human. In certainembodiments, the cell is in a subject in need of treatment.

In certain embodiments, provided is a method of altering transcriptionin a cell which comprises contacting a cell with an effective amount ofa compound of Formula (I), or a pharmaceutically acceptable saltthereof. In certain embodiments, the cell in culture in vitro. Incertain embodiments, the cell is in an animal, e.g., a human. In certainembodiments, the cell is in a subject in need of treatment.

In certain embodiments, a method is provided of selecting a therapy fora subject having a disease associated with PRMT5-mediated disorder ormutation comprising the steps of determining the presence ofPRMT5-mediated disorder or gene mutation in the PRMT5 gene or andselecting, based on the presence of PRMT5-mediated disorder a genemutation in the PRMT5 gene a therapy that includes the administration ofa provided compound. In certain embodiments, the disease is cancer.

In certain embodiments, a method of treatment is provided for a subjectin need thereof comprising the steps of determining the presence ofPRMT5-mediated disorder or a gene mutation in the PRMT5 gene andtreating the subject in need thereof, based on the presence of aPRMT5-mediated disorder or gene mutation in the PRMT5 gene with atherapy that includes the administration of a provided compound. Incertain embodiments, the subject is a cancer patient.

In some embodiments, a provided compound is useful in treating aproliferative disorder, such as cancer, a benign neoplasm, an autoimmunedisease, or an inflammatory disease.

For example, while not being bound to any particular mechanism, PRMT5has been shown to be involved in cyclin D1 dysregulated cancers.Increased PRMT5 activity mediates key events associated with cyclinD1-dependent neoplastic growth including CUL4 repression, CDT1overexpression, and DNA re-replication. Further, human cancers harboringmutations in Fbx4, the cyclin D1 E3 ligase, exhibit nuclear cyclin D1accumulation and increased PRMT5 activity (Aggarwal et al., Cancer Cell.2010 18(4):329-40). Additionally, PRMT5 has also been implicated inaccelerating cell cycle progression through G1 phase and modulatingregulators of G1; for example, PRMT5 may upregulate cyclin-dependentkinase (CDK) 4, CDK6, and cyclins D1, D2 and E1. Moreover, PRMT5 mayactivate phosphoinositide 3-kinase (PI3K)/AKT signaling (Wei et al.,Cancer Sci. 2012 103(9):1640-50). Thus in some embodiments, theinhibition of PRMT5 by a provided compound is useful in treating thefollowing non-limiting list of cancers: breast cancer, esophagealcancer, bladder cancer, lung cancer, hematopoietic cancer, lymphoma,medulloblastoma, rectum adenocarcinoma, colon adenocarcinoma, gastriccancer, pancreatic cancer, liver cancer, adenoid cystic carcinoma, lungadenocarcinoma, head and neck squamous cell carcinoma, brain tumors,hepatocellular carcinoma, renal cell carcinoma, melanoma,oligodendroglioma, ovarian clear cell carcinoma, and ovarian serouscystadenocarcinoma.

In some embodiments, the inhibition of PRMT5 by a provided compound isuseful in treating prostate cancer and lung cancer, in which PRMT5 hasbeen shown to play a role (Gu et al., PLoS One 2012; 7(8):e44033; Gu etal., Biochem. J. (2012) 446 (235-241)). In some embodiments, a providedcompound is useful to delay the onset of, slow the progression of, orameliorate the symptoms of cancer. In some embodiments, a providedcompound is administered in combination with other compounds, drugs, ortherapeutics to treat cancer.

In some embodiments, compounds described herein are useful for treatinga cancer including, but not limited to, acoustic neuroma,adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g.,lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma),appendix cancer, benign monoclonal gammopathy, biliary cancer (e.g.,cholangiocarcinoma), bladder cancer, breast cancer (e.g., adenocarcinomaof the breast, papillary carcinoma of the breast, mammary cancer,medullary carcinoma of the breast), brain cancer (e.g., meningioma;glioma, e.g., astrocytoma, oligodendroglioma; medulloblastoma), bronchuscancer, carcinoid tumor, cervical cancer (e.g., cervicaladenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma,colorectal cancer (e.g., colon cancer, rectal cancer, colorectaladenocarcinoma), epithelial carcinoma, ependymoma, endotheliosarcoma(e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma),endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophagealcancer (e.g., adenocarcinoma of the esophagus, Barrett's adenocarinoma),Ewing sarcoma, eye cancer (e.g., intraocular melanoma, retinoblastoma),familiar hypereosinophilia, gall bladder cancer, gastric cancer (e.g.,stomach adenocarcinoma), gastrointestinal stromal tumor (GIST), head andneck cancer (e.g., head and neck squamous cell carcinoma, oral cancer(e.g., oral squamous cell carcinoma (OS^(cc)), throat cancer (e.g.,laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer,oropharyngeal cancer)), hematopoietic cancers (e.g., leukemia such asacute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acutemyelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronicmyelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chroniclymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL); lymphoma suchas Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkinlymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma(DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)), follicularlymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma(CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas(e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodalmarginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma),primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacyticlymphoma (i.e., “Waldenstrim's macroglobulinemia”), hairy cell leukemia(HCL), immunoblastic large cell lymphoma, precursor B-lymphoblasticlymphoma and primary central nervous system (CNS) lymphoma; and T-cellNHL such as precursor T-lymphoblastic lymphoma/leukemia, peripheralT-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g.,mycosis fungiodes, Sezary syndrome), angioimmunoblastic T-cell lymphoma,extranodal natural killer T-cell lymphoma, enteropathy type T-celllymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplasticlarge cell lymphoma); a mixture of one or more leukemia/lymphoma asdescribed above; and multiple myeloma (MM)), heavy chain disease (e.g.,alpha chain disease, gamma chain disease, mu chain disease),hemangioblastoma, inflammatory myofibroblastic tumors, immunocyticamyloidosis, kidney cancer (e.g., nephroblastoma a.k.a. Wilms' tumor,renal cell carcinoma), liver cancer (e.g., hepatocellular cancer(H^(cc)), malignant hepatoma), lung cancer (e.g., bronchogeniccarcinoma, small cell lung cancer (SCLC), non-small cell lung cancer(NSCLC), adenocarcinoma of the lung), leiomyosarcoma (LMS), mastocytosis(e.g., systemic mastocytosis), myelodysplastic syndrome (MDS),mesothelioma, myeloproliferative disorder (MPD) (e.g., polycythemia Vera(PV), essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM)a.k.a. myelofibrosis (MF), chronic idiopathic myelofibrosis, chronicmyelocytic leukemia (CML), chronic neutrophilic leukemia (CNL),hypereosinophilic syndrome (HES)), neuroblastoma, neurofibroma (e.g.,neurofibromatosis (NF) type 1 or type 2, schwannomatosis),neuroendocrine cancer (e.g., gastroenteropancreatic neuroendoctrinetumor (GEP-NET), carcinoid tumor), osteosarcoma, ovarian cancer (e.g.,cystadenocarcinoma, ovarian embryonal carcinoma, ovarianadenocarcinoma), papillary adenocarcinoma, pancreatic cancer (e.g.,pancreatic andenocarcinoma, intraductal papillary mucinous neoplasm(IPMN), Islet cell tumors), penile cancer (e.g., Paget's disease of thepenis and scrotum), pinealoma, primitive neuroectodermal tumor (PNT),prostate cancer (e.g., prostate adenocarcinoma), rectal cancer,rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g., squamouscell carcinoma (S^(cc)), keratoacanthoma (KA), melanoma, basal cellcarcinoma (B^(cc))), small bowel cancer (e.g., appendix cancer), softtissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma,malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma,fibrosarcoma, myxosarcoma), sebaceous gland carcinoma, sweat glandcarcinoma, synovioma, testicular cancer (e.g., seminoma, testicularembryonal carcinoma), thyroid cancer (e.g., papillary carcinoma of thethyroid, papillary thyroid carcinoma (PTC), medullary thyroid cancer),urethral cancer, vaginal cancer and vulvar cancer (e.g., Paget's diseaseof the vulva).

In some embodiments, a provided compound is useful in treating ametabolic disorder, such as diabetes or obesity. For example, while notbeing bound to any particular mechanism, a role for PRMT5 has beenrecognized in adipogenesis. Inhibition of PRMT5 expression in multiplecell culture models for adipogenesis prevented the activation ofadipogenic genes, while overexpression of PRMT5 enhanced adipogenic geneexpression and differentiation (LeBlanc et al., Mol Endocrinol. 2012April; 26(4):583-97). Additionally, it has been shown that adipogenesisplays a pivotal role in the etiology and progression of diabetes andobesity (Camp et al., Trends Mol Med. 2002 September;8(9):442-7). Thusin some embodiments, the inhibition of PRMT5 by a provided compound isuseful in treating diabetes and/or obesity.

In some embodiments, a provided compound is useful to delay the onsetof, slow the progression of, or ameliorate the symptoms of, diabetes. Insome embodiments, the diabetes is Type 1 diabetes. In some embodiments,the diabetes is Type 2 diabetes. In some embodiments, a providedcompound is useful to delay the onset of, slow the progression of, orameliorate the symptoms of, obesity. In some embodiments, a providedcompound is useful to make a subject lose weight. In some embodiments, aprovided compound could be used in combination with other compounds,drugs, or therapeutics, such as metformin and insulin, to treat diabetesand/or obesity.

In some embodiments, a provided compound is useful in treating a blooddisorder, e.g., a hemoglobinopathy, such as sickle cell disease orβ-thalassemia. For example, while not being bound to any particularmechanism, PRMT5 is a known repressor of γ-globin gene expression, andincreased fetal γ-globin (HbF) levels in adulthood are associated withsymptomatic amelioration in sickle cell disease and β-thalassemia (Xu etal., Haematologica. 2012 November; 97(11):1632-40). Thus in someembodiments, the inhibition of PRMT5 by a provided compound is useful intreating a blood disorder, such as a hemoglobinopathy such as sicklecell disease or β-thalassemia.

In some embodiments, a provided compound is useful to delay the onsetof, slow the progression of, or ameliorate the symptoms of, sickle celldisease. In some embodiments, a provided compound is useful to delay theonset of, slow the progression of, or ameliorate the symptoms of,β-thalassemia. In some embodiments, a provided compound could be used incombination with other compounds, drugs, or therapeutics, to treat ahemoglobinopathy such as sickle cell disease or β-thalassemia.

In some embodiments, compounds described herein can prepared usingmethods shown in general Scheme 1 ring opening of a chiral or racemicepoxide group to form an amino alcohol moiety. A ring opening step canbe performed in either direction as shown in scheme 1.

In some embodiments, compounds described herein can prepared usingmethods shown in general Scheme 2. Compound B can be prepared via ringopening of a chiral or racemic epoxide group. This amino alcoholintermediate can be coupled to form an amide via normal amide couplingmethodology using a carboxylic acid A wherein Z₁ is hydrogen or viaamination of an ester of intermediate A when Z₁ is an optionallysubstituted aliphatic group.

In some embodiments, compounds described herein can prepared usingmethods shown in general Scheme 3. Compound B can be prepared via ringopening of a chiral or racemic epoxide group. This amino alcoholintermediate can be coupled to form an amide via normal amide couplingmethodology using a carboxylic acid A wherein Z₁ is hydrogen or viaamination of an ester of intermediate A when Z₁ is an optionallysubstituted aliphatic group

In some embodiments, compounds described herein can prepared usingmethods shown in general Scheme 4, which describes ring opening of achiral or racemic epoxide group to form the amino alcohol moiety linker.

EXAMPLES

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

Synthetic Methods Compound 2tert-butyl(3-((2-hydroxy-3-(isoindolin-2-yl)propyl)carbamoyl)phenyl)carbamate

To a solution of 3-((tert-butoxycarbonyl)amino)benzoic acid (300 mg, 1.3mmol) in DCM (8 mL) was added EDCI (383 mg, 2.0 mmol), HOBt (270 mg, 2.0mmol), Et₃N (263 mg, 2.6 mmol), 1-amino-3-(isoindolin-2-yl)propan-2-ol(499 mg, 2.6 mmol), and the mixture was stirred at 25° C. for 16 h. Thecrude reaction mixture was washed with water and extracted with DCM. Thecombined organic layers were concentrated, and the residue was purifiedby column chromatography to yield the desired product (DCM: MeOH=10:1).(260 mg, yield 49%) MS (ESI⁺) e/z: 412.2 [M+1]⁺. ¹H NMR (MeOD, 400 MHz),δ ppm: 7.87 (s, 1H), 7.58-7.50 (m, 1H), 7.45-7.38 (m, 1H), 7.34-7.28 (m,1H), 7.25-7.14 (m, 4H), 4.08-3.96 (m, 5H), 3.63-3.53 (m, 1H), 3.45-3.37(m, 1H), 2.94-2.80 (m, 2H), 1.53 (s, 9H).

Compound 3(R)—N—((R)-2-hydroxy-3-(isoindolin-2-yl)propyl)-2-(quinolin-8-yloxy)propanamide

To a solution of (R)-methyl 2-(quinolin-8-yloxy)propanoate (100 mg,0.433 mmol) in EtOH (1 mL) was added(R)-1-amino-3-(isoindolin-2-yl)propan-2-ol (83 mg, 0.433 mmol). Thereaction mixture was heated under microwave conditions at 120° C. for0.5 h, concentrated, and purified by preparative TLC first and then bypreparative HPLC purification. (19 mg, yield 11%) MS (ESI⁺) e/z: 392.1[M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 8.90 (d, J=2.76 Hz, 1H), 8.34 (d,J=8.28 Hz, 1H), 7.64-7.48 (m, 3H), 7.34-7.24 (m, 1H), 7.23-7.08 (m, 4H),5.07 (q, J=6.53 Hz, 1H), 3.97-3.71 (m, 5H), 3.49 (dd, J=13.55, 5.27 Hz,1H), 3.29 (d, J=6.27 Hz, 1H), 2.71 (d, J=6.02 Hz, 2H), 1.72 (d, J=6.53Hz, 3H).

Compound 4(S)—N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-2-phenoxyacetamide

Step 1: (R)-2-(oxiran-2-ylmethyl)isoindoline

To a solution of isoindoline (500 mg, 4.20 mmol) and(S)-oxiran-2-ylmethyl 3-nitrobenzenesulfonate (1.27 g, 5.04 mmol) in THF(100 mL) was added KF (580 mg, 10 mmol) at 0° C. The reaction mixturewas warmed to 25° C., stirred for 16 h, filtered and concentrated. Thecrude product was used in the next step without further purification.(600 mg, yield 68%) MS (ESI⁺) e/z: 176.1 [M+1]⁺.

Step 2: (S)-1-amino-3-(isoindolin-2-yl)propan-2-ol

EtOH (50 mL) was cooled to −78° C. and ammonia gas was bubbled throughthe solution. To the solution was added(R)-2-(oxiran-2-ylmethyl)isoindoline (280 mg, 1.6 mmol), the reactionvessel was sealed and heated at 80° C. for 4 h. The reaction mixture wascooled, concentrated and the crude product was used in the next stepwithout further purification. (600 mg, yield 91%) MS (ESI⁺) e/z: 1193.1[M+1]⁺.

Step 3: (S)—N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-2-phenoxyacetamide

A solution of (S)-1-amino-3-(isoindolin-2-yl)propan-2-ol (100 mg, 0.52mmol), ethyl 2-(quinolin-8-yloxy)acetate (120 mg, 0.52 mmol) in EtOH (1mL) was heated under microwave conditions at 120° C. for 0.5 h. Thereaction mixture was concentrated and purified by preparative HPLCpurification. (60 mg, yield 31%) MS (ESI⁺) e/z: 378.1 [M+1]⁺. ¹H NMR(MeOD, 400 MHz), δ ppm: 8.93-8.88 (m, 1H), 8.44-8.38 (m, 1H), 7.66-7.57(m, 3H), 7.34-7.29 (m, 1H), 7.24-7.14 (m, 4H), 4.82-4.79 (m, 2H),4.08-4.03 (m, 1H), 3.99 (s, 4H), 3.60-3.55 (m, 1H), 3.46-3.40 (m, 1H),2.91-2.85 (m, 1H), 2.8-2.78 (m, 1H).

Compound 5(R)—N—((S)-2-hydroxy-3-(isoindolin-2-yl)propyl)-2-(quinolin-8-yloxy)propanamide

Step 1: (R)-methyl 2-(quinolin-8-yloxy)propanoate

To a solution of quinolin-8-ol (300 mg, 2.07 mmol) in THF (5 mL) wasadded (S)-methyl 2-hydroxypropanoate (215 mg, 2.07 mmol), PPh₃ (647 mg,2.47 mmol), DEAD (430 mg, 2.47 mmol) and the resulting mixture wasstirred at 25° C. for 16 h. Aqueous HCl (1M, 10 mL) was added, themixture separated, and the aqueous layer extracted with ethyl acetate.The aqueous portion was basified by addition of saturated aqueous NaHCO₃(20 mL) and extracted with ethyl acetate. The combined organic extractswere washed with brine, dried over sodium sulfate and concentrated. Theresidue was purified by column chromatography. (300 mg, yield 63%) MS(ESI⁺) e/z: 232.1 [M+1]⁺.

(R)—N—((S)-2-hydroxy-3-(isoindolin-2-yl)propyl)-2-(quinolin-8-yloxy)propanamide

To a solution of (R)-methyl 2-(quinolin-8-yloxy)propanoate (100 mg,0.433 mmol) in EtOH (1 mL) was added(S)-1-amino-3-(isoindolin-2-yl)propan-2-ol (83 mg, 0.433 mmol). Thereaction mixture was heated under microwave conditions at 120° C. for0.5 h. The mixture was concentrated and purified by preparative HPLCpurification. (15 mg, yield 8.8%) MS (ESI⁺) e/z: 392.1 [M+1]⁺. ¹H NMR(MeOD, 400 MHz), δ ppm: ¹H NMR (400 MHz, METHANOL-d₄) δ 8.90 (dd,J=4.27, 1.76 Hz, 1H) 8.33 (dd, J=8.41, 1.63 Hz, 1H) 7.51-7.64 (m, 3H)7.29 (dd, J=6.15, 2.64 Hz, 1H) 7.08-7.22 (m, 4H) 5.08 (q, J=6.78 Hz, 1H)3.75-3.93 (m, 5H) 3.36-3.43 (m, 2H) 2.55-2.66 (m, 2H) 1.73 (d, J=6.78Hz, 3H).

Compound 6N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-3-((tetrahydro-2H-pyran-4-yl)amino)benzamide

To a solution of3-amino-N-(2-hydroxy-3-(isoindolin-2-yl)propyl)benzamide (120 mg, 0.39mmol) in MeOH (10 mL) was added dihydro-2H-pyran-4(3H)-one (78 mg, 0.78mmol) and AcOH (0.05 mL). The mixture was stirred at 25° C. for 2 h.NaBH₃CN (123 mg, 1.95 mmol) was added, and the resulting mixture wasstirred at 25° C. for 16 h. The reaction mixture was concentrated, andthe residue was dissolved in water, extracted with ethyl acetate, driedover sodium sulfate and concentrated. The crude product was purified bypreparative HPLC purification. (22 mg, yield 14%) MS (ESI⁺) e/z: 396.2[M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 7.62-7.53 (m, 2H), 7.53-7.36 (m,5H), 7.30-7.21 (m, 1H), 4.76-4.57 (m, 2H), 4.34-4.24 (m, 1H), 4.08-3.96(m, 2H), 3.74-3.65 (m, 1H), 3.65-3.57 (m, 2H), 3.57-3.43 (m, 4H),3.36-3.32 (m, 2H), 2.03-1.91 (m, 2H), 1.72-1.57 (m, 2H).

Compound 82-(2-(1H-pyrazol-3-yl)phenoxy)-N-(2-hydroxy-3-(isoindolin-2-yl)propyl)acetamide

Step 1: ethyl 2-(2-(1H-pyrazol-3-yl)phenoxy)acetate

To a solution of 2-(1H-pyrazol-3-yl) phenol (400 mg, 2.5 mmol) and K₂CO₃(518 mg, 3.75 mmol) in CH₃CN (10 mL) was added ethyl 2-bromoacetate (418g, 2.5 mmol). The mixture was stirred at room temperature for 2 h,diluted with water, extracted with ethyl acetate and concentrated. Thecrude material was purified by column chromatography. (129 mg, yield21%) MS (ESI⁺) e/z: 247.2 [M+1]⁺.

Step 2:2-(2-(1H-pyrazol-3-yl)phenoxy)-N-(2-hydroxy-3-(isoindolin-2-yl)propyl)acetamide

Ethyl 2-(2-(1H-pyrazol-3-yl)phenoxy)acetate (129 mg, 0.52 mmol) and1-amino-3-(isoindolin-2-yl)propan-2-ol (100 mg, 0.52 mmol) weredissolved in EtOH (1 mL) and heated under microwave conditions at 120°C. for 1 h. The reaction mixture was concentrated and purified bypreparative HPLC purification. (16.2 mg, yield 8%) MS (ESI⁺) e/z: 393.2[M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 7.70 (d, J=7.53 Hz, 2H),7.39-7.32 (m, 1H), 7.26-7.14 (m, 4H), 7.14-7.05 (m, 2H), 6.74 (d, J=1.76Hz, 1H), 4.70 (br. s., 2H), 3.97 (s, 5H), 3.54 (dd, J=13.55, 4.77 Hz,1H), 3.38 (d, J=8.53 Hz, 1H), 2.81 (d, J=4.27 Hz, 2H).

Compound 91-(isoindolin-2-yl)-3-(3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

Step 1: 5-bromo-N-methyl-2-nitroaniline

To a solution of 4-bromo-2-fluoro-1-nitrobenzene (10 g, 45.7 mmol) inDMSO (50 mL) was added TEA (18.47 g, 183 mmol), methylaminehydrochloride (6.1 g, 91.4 mmol) and the reaction mixture was heatedunder microwave conditions at 120° C. for 3 h. The mixture was cooled,extracted with ethyl acetate; the combined organic extracts were washedwith brine, dried over sodium sulfate and concentrated. The crudeproduct was used in next step without further purification. (10.5 g,yield 98%) MS (ESI⁺) e/z: 231.1.

Step 2: 5-bromo-N1-methylbenzene-1,2-diamine

To a solution of 5-bromo-N-methyl-2-nitroaniline (10 g, 43.5 mmol) inEtOH/H₂O (700 mL) was added Fe (14.6 g, 261 mmol), ammonium chloride (14g, 261 mmol) and the reaction mixture was heated at 60° C. for 4 h. Thecrude reaction mixture was filtered, concentrated, dissolved in ethylacetate, washed with brine, dried over sodium sulfate, filtered andconcentrated. The crude product was used in next reaction withoutfurther purification. (7.9 g, yield 90%) MS (ESI⁺) e/z: 202.1.

Step 3: 6-bromo-1-methyl-1H-benzo[d]imidazole

To a solution of 5-bromo-N1-methylbenzene-1,2-diamine (7.4 g, 37 mmol)in trimethyl orthoformate (100 mL) was added p-toluenesulfonic acid(0.36 g, 1.9 mmol). The reaction mixture was heated at 100° C. for 4 h,cooled, concentrated, dissolved in ethyl acetate, washed with brine,dried over sodium sulfate and concentrated. The crude product was usedin next step without further purification. (7.3 g, yield 93%) MS (ESI⁺)e/z: 212.1.

Step 4:1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole

To a solution of 6-bromo-1-methyl-1H-benzo[d]imidazole (5 g, 23.8 mmol)in dioxane (60 mL) was added4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (9.1 g, 35.7mmol), Pd(dppf)Cl₂ (0.5 g) and potassium acetate (4.67 g, 47.6 mmol).The reaction mixture was heated at 100° C. for 3 h, cooled andconcentrated. The crude reaction mixture was purified by columnchromatography. (6 g, yield 98%) MS (ESI⁺) e/z: 259.1.

Step 5: 3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenol

To a solution of1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazole(6 g, 23.1 mmol) in dioxane/water (50 mL) was added 3-bromophenol (4.8g, 27.7 mmol), Pd(dppf)Cl₂ (0.3 g) and Cs₂CO₃ (15 g, 46.2 mmol). Thereaction mixture was heated at 100° C. for 3 h, cooled and concentrated.The crude reaction mixture was purified by column chromatography. (4.8g, yield 92%) MS (ESI⁺) e/z: 225.1.

Step 6: 1-methyl-6-(3-(oxiran-2-ylmethoxy)phenyl)-1H-benzo[d]imidazole

To a solution of NaH (161 mg, 6.69 mmol) in DMF (5 mL) was added3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenol (500 mg, 2.23 mmol) at 27°C. After 0.5 h, 2-(chloromethyl)oxirane (246 mg, 2.68 mmol) was addedand the reaction mixture was stirred 16 h, diluted with water andextracted with ethyl acetate. The combined organic extracts were washedwith brine, dried over sodium sulfate, filtered and concentrated. Thecrude product was used in next step without further purification. (500mg, yield 80%).

Step 7:1-(isoindolin-2-yl)-3-(3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

To a solution of1-methyl-6-(3-(oxiran-2-ylmethoxy)phenyl)-1H-benzo[d]imidazole (500 mg,1.78 mmol) in MeOH (5 mL) was added isoindoline (213 mg, 1.78 mmol) at25° C., and the mixture was heated to reflux and stirred for 16 h. Thereaction mixture was cooled, concentrated and purified by preparativeHPLC purification. (80 mg, yield 11%) MS (ESI⁺) e/z: 400.2 [M+1]⁺. ¹HNMR (MeOD, 400 MHz), δ ppm: 8.22 (s, 1H), 7.87 (s, 1H), 7.71 (d, J=8.4Hz, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.41-7.19 (m, 7H), 6.97-6.95 (m, 1H),4.16-4.04 (m, 7H), 3.89 (s, 3H), 3.04-2.87 (m, 2H).

Compound 10N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-2-((4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yl)oxy)acetamide

Step 1: 2-aminobenzene-1,3-diol

To a solution of 2-nitrobenzene-1,3-diol (5.00 g, 32.2 mmol) in MeOH(100 mL) was added Pd/C (200 mg). The mixture was stirred under a H₂atmosphere at 25° C. for 16 h, filtered, and concentrated. The crudeproduct was used in the next step without further purification. (3.00 g,yield 75%).

Step 2: 5-hydroxy-2H-benzo[b][1,4]oxazin-3(4H)-one

To a solution of 2-aminobenzene-1,3-diol (2.00 g, 16.0 mmol) and TEA(1.94 g, 19.2 mmol) in anhydrous DMF (30 mL) was added 2-chloroacetylchloride (1.81 g, 16.0 mmol). After 16 h, K₂CO₃ (2.65 g, 19.2 mmol) wasadded and the reaction mixture was stirred for another 16 h. The mixturewas diluted with water and extracted with DCM. The combined organicextracts were washed with water, brine, the combined organic extractsdried over sodium sulfate, filtered and concentrated. The crude productwas purified by column chromatography. (1.7 g, yield 65%).

Step 3: ethyl2-(3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)acetate

A solution of 5-hydroxy-2H-benzo[b][1,4]oxazin-3(4H)-one (100 mg, 0.604mmol) and K₂CO₃ (167 mg, 1.21 mmol) in anhydrous DMF (5 mL) was stirredat 27° C. for 5 minutes, then ethyl 2-bromoacetate (121 mg, 0.727 mmol)was added. The reaction mixture was stirred at 27° C. for 16 h andconcentrated. MS (ESI⁺) e/z: 238.0 [M+1]⁺.

Step 4: ethyl2-(4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)acetate

To the solution of ethyl2-(3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy) acetate (72 mg,0.287 mmol) and K₂CO₃ (39.6 mg, 0.287 mmol) in DMF (5 mL) was added MeI(40.7 mg, 0.287 mmol). The mixture was stirred at 25° C. for 16 h,partitioned between water (50 mL) and DCM (100 mL), the organic portionwas washed with water, brine, dried over sodium sulfate, filtered andconcentrated. The crude product was purified by preparative TLC. (43 mg,yield 57%). ¹H NMR (MeOD, 400 MHz), δ ppm: 6.99 (t, J=8.3 Hz, 1H), 6.74(d, J=7.5 Hz, 1H), 6.55 (d, J=8.3 Hz, 1H), 4.69 (s, 2H), 4.51 (s, 2H),4.30 (q, J=7.2 Hz, 2H), 3.56 (s, 3H), 1.34-1.33 (m, 1H), 1.33 (t, J=7.2Hz, 3H).

Step 5: ethyl2-((4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yl)oxy)acetate

A solution of ethyl2-((4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yl)oxy)acetate(100 mg, 0.377 mmol) in anhydrous THF (3 mL) was cooled to 0° C. andBH₃-Me₂S (0.1 mL) added. The solution was stirred at 27° C. for 6 h,diluted with methanol and concentrated. The residue was partitionedbetween ethyl acetate (30 mL) and water (20 mL), the organic layerwashed with brine, dried over sodium sulfate, filtered and concentrated.The crude product was purified by preparative TLC. (72 mg, yield 76%) MS(ESI⁺) e/z: 252.2 [M+1]⁺. ¹H NMR (CDCl₃, 400 MHz), δ ppm: 6.87-6.76 (m,1H), 6.62-6.52 (m, 1H), 6.40-6.28 (m, 1H), 4.70 (s, 2H), 4.34-4.22 (m,2H), 4.18-4.06 (m, 2H), 3.21-3.07 (m, 2H), 2.99-2.85 (m, 3H), 1.33-1.25(m, 3H).

Step 6:N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-2-((4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yl)oxy)acetamide

A solution of ethyl2-((4-methyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yl)oxy) acetate (72mg, 0.287 mmol), 1-amino-3-(isoindolin-2-yl)ropan-2-ol (55 mg, 0.755mmol) in EtOH (0.1 mL) was heated under microwave conditions at 120° C.for 2 h. The reaction mixture was concentrated and purified bypreparative HPLC purification. (17.4 mg, yield 15%). MS (ESI⁺) e/z:398.2 [M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 7.27-7.13 (m, 4H), 6.91 (t,J=8.3 Hz, 1H), 6.66-6.50 (m, 2H), 4.66-4.59 (m, 2H), 4.23-4.09 (m, 2H),4.02-3.86 (m, 5H), 3.49 (dd, J=4.9, 13.7 Hz, 1H), 3.32-3.27 (m, 1H),3.21-3.11 (m, 2H), 2.92-2.81 (m, 3H), 2.80-2.70 (m, 2H).

Compound 11tert-butyl(3-((2-hydroxy-3-(isoindolin-2-yl)propyl)carbamoyl)phenyl)carbamate

Step 1: 3-((tert-butoxycarbonyl)amino)benzoic acid

To a solution of 3-aminobenzoic acid (1.37 g, 10 mmol) in THF (20 mL)and H₂O (2 mL) was added Boc₂O (2.18 g, 10 mmol) and Et₃N (1.52 g, 15mmol). The reaction mixture was stirred at 25° C. for 16 h,concentrated, dissolved in water and extracted with ethyl acetate. Thecombined extracts were concentrated and the crude product was used inthe next step without further purification. (2.5 g, yield 96%) MS (ESI⁺)e/z: 260.0 [M+1]⁺.

Step 2:tert-butyl(3-((2-hydroxy-3-(isoindolin-2-yl)propyl)carbamoyl)phenyl)carbamate

To a solution of 3-((tert-butoxycarbonyl)amino)benzoic acid (300 mg, 1.3mmol) in DCM (8 mL) was added EDCI (383 mg, 2.0 mmol), HOBt (270 mg, 2.0mmol), Et₃N (263 mg, 2.6 mmol), 1-amino-3-(isoindolin-2-yl)propan-2-ol(499 mg, 2.6 mmol), and the mixture was stirred at 25° C. for 16 h. Thecrude reaction mixture was washed with water and extracted with DCM. Thecombined organic layers were concentrated, and the residue was purifiedby column chromatography (DCM: MeOH=10:1). (260 mg, yield 49%) MS (ESI⁺)e/z: 412.2 [M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 7.87 (s, 1H),7.58-7.50 (m, 1H), 7.45-7.38 (m, 1H), 7.34-7.28 (m, 1H), 7.25-7.14 (m,4H), 4.08-3.96 (m, 5H), 3.63-3.53 (m, 1H), 3.45-3.37 (m, 1H), 2.94-2.80(m, 2H), 1.53 (s, 9H).

Compound 12N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-2-(2-(methylsulfonyl)phenoxy)acetamide

Step 1: ethyl 2-(2-(methylsulfonyl)phenoxy)acetate

To a solution of 2-(methylsulfonyl)phenol (100 mg, 0.58 mmol) and K₂CO₃(276 mg, 2 mmol) in CH₃CN (10 mL) was added ethyl 2-bromoacetate (115.2mg, 0.69 mmol) at 25° C. The mixture was heated at 80° C. for 4 h,diluted with water and extracted with ethyl acetate. The combinedorganic layers were washed with brine, dried over sodium sulfate andconcentrated. The crude product was used in the next step withoutfurther purification. (140 mg, yield 94%).

Step 2:N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-2-(2-(methylsulfonyl)phenoxy)acetamide

Ethyl 2-(2-(methylsulfonyl)phenoxy)acetate (50 mg, 0.19 mmol) and1-amino-3-(isoindolin-2-yl)propan-2-ol (36 mg, 0.19 mmol) were dissolvedin EtOH (1 mL) and heated under microwave conditions at 120° C. 0.5 h.The reaction mixture was concentrated and purified by preparative HPLCpurification. (11 mg, yield 14%) MS (ESI⁺) e/z: 405.2 [M+1]⁺. ¹H NMR(MeOD, 400 MHz), δ ppm: 7.98-7.91 (m, 1H), 7.76-7.69 (m, 1H), 7.33-7.24(m, 2H), 7.21 (d, J=2.76 Hz, 4H), 4.86-4.80 (m, 2H), 4.01-3.97 (m, 4H),3.97-3.91 (m, 1H), 3.57-3.50 (m, 1H), 3.39-3.35 (m, 1H), 3.30 (s, 3H),2.86-2.81 (m, 1H), 2.81-2.74 (m, 1H).

Compound 13N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-3-(pyridin-2-yl)benzamide

Step 1: methyl 3-(pyridin-2-yl)benzoate

A solution (3-(methoxycarbonyl)phenyl)boronic acid (500 mg, 2.78 mmol),2-bromopyridine (399 mg, 2.53 mmol), K₂CO₃ (1.0 g, 7.6 mmol) andPd(dppf)Cl₂ (20 mg) in a mixture of dioxane (10 mL) and water (2.5 mL)was heated under microwave conditions at 120° C. for 0.5 h. The reactionmixture was filtered, concentrated, and the crude product was purifiedby column chromatography eluting with petroleum ether/ethyl acetate(5:1). (400 mg, yield 74%) MS (ESI⁺) e/z: 214.1 [M+1]⁺.

Step 2: 3-(pyridin-2-yl)benzoic acid

To a solution of methyl 3-(pyridin-2-yl)benzoate (400 mg, 1.88 mmol) inMeOH (3 mL) was added aqueous of NaOH (1 mL, 40 mol %). The reactionmixture was stirred at room temperature for 3 h and concentrated. Thecrude residue was dissolved in water and the pH was adjusted to 5˜6 with2N of HCl. The solution was extracted with ethyl acetate, brine, driedover sodium sulfate, filtered and concentrated. (350 mg, yield 93%) MS(ESI⁺) e/z: 200.1 [M+1]⁺.

Step 3:N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-3-(pyridin-2-yl)benzamide

To a solution of 3-(pyridin-2-yl)benzoic acid (60 mg, 0.30 mmol) in DCM(8 mL) was added EDCI (86 mg, 0.45 mmol), HOBt (61 mg, 0.45 mmol), Et₃N(61 mg, 0.6 mmol) and 1-amino-3-(isoindolin-2-yl)propan-2-ol (86 mg,0.45 mmol). The mixture was stirred at 25° C. for 16 h, washed withwater and extracted with DCM. The combined organic extracts wereconcentrated, and the residue was purified by preparative HPLCpurification. (30 mg, 27%) MS (ESI⁺) e/z: 374.2 [M+1]⁺. ¹H NMR (MeOD,400 MHz), δ ppm: 8.65 (d, J=4.8 Hz, 1H), 8.15 (d, J=8.0 Hz, 1H),7.96-7.92 (m, 3H), 7.62 (dd, J=7.2 Hz, 1H), 7.40-7.35 (m, 6H), 4.70 (s,4H), 4.29 (br.s, 1H), 3.63-3.30 (m, 4H).

Compound 152-(2-(N,N-dimethylsulfamoyl)phenoxy)-N-(2-hydroxy-3-(isoindolin-2-yl)propyl)acetamide

Step 1: ethyl 2-(2-bromophenoxy)acetate

To a solution of 2-bromophenol (2 g, 0.0116 mol) in MeCN (10 mL) wasadded ethyl bromoacetate (2.12 g, 0.0128 mol) and K₂CO₃ (4.81 g, 0.035mol). The mixture was stirred at 80° C. for 4 h, filtered andconcentrated. The crude product was used in the next step withoutfurther purification.

Step 2: ethyl 2-(2-((4-methoxybenzyl)thio)phenoxy)acetate

To a solution of ethyl 2-(2-bromophenoxy)acetate (3.0 g, 0.0116 mol) indioxane (30 mL) was added (4-methoxyphenyl)methanethiol (2.14 g, 0.0139mol), Pd₂(dba)₃ (20 mg), xantphos (20 mg) and DIEA (3 mL). The mixturewas degassed 4 times (N₂) and heated at reflux for 3 h, concentrated,and the crude product was purified by column chromatography. (3 g, yield79%) MS (ESI⁺) e/z: 355.1 [M+1]⁺.

Step 3: ethyl 2-(2-(chlorosulfonyl)phenoxy)acetate

To a solution of ethyl 2-(2-((4-methoxybenzyl)thio)phenoxy)acetate (1.0g, 3.01 mmol) in MeCN:HOAC:H₂O (80:1:2, 10 mL) was added1,3-dichloro-5,5-dimethylhydantoin (1.19 g, 6.02 mmol) at 0° C. Themixture was stirred at 0° C. for 3 h and concentrated. The crude residuewas extracted with DCM, aqueous NaHCO₃ that had been cooled to 10° C.,the combined organic extracts were washed with brine, dried overanhydrous sodium sulfate and concentrated. The crude product was used inthe next step without further purification.

Step 4: ethyl 2-(2-(N,N-dimethylsulfamoyl)phenoxy acetate

To a solution of ethyl 2-(2-(chlorosulfonyl)phenoxy)acetate (300 mg,1.08 mmol) in pyridine (5 mL) was added dimethylamine hydrochloride (105mg, 1.30 mmol) at 0° C. The mixture was warmed to 25° C. and stirred for16 h. The reaction mixture was concentrated and the crude product wasused in the next step without further purification. (250 mg, yield 81%)MS (ESI⁺) e/z: 288.0 [M+1]⁺.

Step 5:2-(2-(N,N-dimethylsulfamoyl)phenoxy)-N-(2-hydroxy-3-(isoindolin-2-yl)propyl)acetamide

To a solution of ethyl 2-(2-(N,N-dimethylsulfamoyl)phenoxy)acetate (100mg, 0.35 mmol) in EtOH (1 mL) was added1-amino-3-(isoindolin-2-yl)propan-2-ol (66.8 mg, 0.35 mmol). Thereaction mixture was heated under microwave conditions at 120° C. for0.5 h. The material was concentrated and purified by preparative HPLCpurification. (20 mg, yield 13%) MS (ESI⁺) e/z: 434.1 [M+1]⁺. ¹H NMR(MeOD, 400 MHz), δ ppm: 7.85 (dd, J=7.78, 1.51 Hz, 1H), 7.70-7.61 (m,1H), 7.27-7.16 (m, 6H), 4.75 (s, 2H), 4.05-3.91 (m, 5H), 3.55 (dd,J=13.68, 4.89 Hz, 1H), 3.40-3.35 (m, 1H), 2.92-2.77 (m, 8H).

Compound 16(R)—N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-2-(quinolin-8-yloxy)acetamide

Step 1: ethyl 2-(quinolin-8-yloxy)acetate

To a solution of quinolin-8-ol (3 g, 0.0207 mol) in MeCN (10 mL) wasadded ethyl bromoacetate (4.12 g, 0.025 mol) and K₂CO₃ (5.75 g, 0.0414mol). The mixture was stirred at 80° C. for 12 h, filtered andconcentrated. The crude residue was purified by column chromatography.(3.9 g, yield, 82%) MS (ESI⁺) e/z: 232.1 [M+1]⁺.

Step 2:(R)—N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-2-(quinolin-8-yloxy)acetamide

(R)-1-amino-3-(isoindolin-2-yl)propan-2-ol (100 mg, 0.52 mmol) and ethyl2-(quinolin-8-yloxy)acetate (120 mg, 0.52 mmol) were dissolved in EtOH(1 mL) and heated under microwave conditions at 120° C. for 0.5 h. Thereaction mixture was concentrated and purified by preparative HPLCpurification. (100 mg, yield 51%) MS (ESI⁺) e/z: 378.1 [M+1]⁺. ¹H NMR(MeOD, 400 MHz), δ ppm: 8.95-8.85 (m, 1H), 8.44-8.34 (m, 1H), 7.69-7.54(m, 3H), 7.33-7.27 (m, 1H), 7.18 (s, 4H), 4.79 (s, 2H), 4.09-4.03 (m,1H), 3.99 (s, 4H), 3.61-3.53 (m, 1H), 3.47-3.38 (m, 1H), 2.91-2.85 (m,1H), 2.84-2.77 (m, 1H).

Compound 17(R)-1-(isoindolin-2-yl)-3-(3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

Step 1:(S)-3-(1-methyl-1H-benzo[d]imidazol-6-yl)-N-(oxiran-2-ylmethyl)aniline

To a solution of NaH (161 mg, 6.69 mmol) in DMF (5 mL) was added3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenol (500 mg, 2.23 mmol) at 27°C. After 0.5 h, (S)-2-(chloromethyl)oxirane (246 mg, 2.68 mmol) wasadded and the reaction mixture was stirred 16 h, diluted with water andextracted with ethyl acetate. The combined organic extracts were washedwith brine, dried over sodium sulfate, filtered and concentrated. Thecrude product was used in next step without further purification. (480mg, yield 77%).

Step 2:(R)-1-(isoindolin-2-yl)-3-(3-(1-methyl-1H-benzo[d]imidazol-6-yl)phenoxy)propan-2-ol

To a solution of(S)-3-(1-methyl-1H-benzo[d]imidazol-6-yl)-N-(oxiran-2-ylmethyl) aniline(480 mg, 1.71 mmol) in MeOH (5 mL) was added isoindoline (213 mg, 1.78mmol) at 25° C., and the mixture was heated to reflux and stirred for 16h. The reaction mixture was cooled, concentrated and purified bypreparative HPLC and SFC purification. (104.8 mg, yield 15%) MS (ESI⁺)e/z: 400.2 [M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 8.44-8.26 (m, 1H),8.17 (s, 1H), 7.82-7.66 (m, 2H), 7.63-7.52 (m, 1H), 7.44-7.37 (m, 5H),7.36-7.28 (m, 2H), 6.99 (dd, J=8.03, 1.63 Hz, 1H), 4.78 (s, 4H), 4.47(dd, J=8.97, 3.58 Hz, 1H), 4.23-4.10 (m, 2H), 3.94 (s, 3H), 3.74-3.58(m, 2H).

Compound 19N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-2-(quinolin-8-yloxy)acetamide

Step 1: 2-(oxiran-2-ylmethyl)isoindoline

To a solution of isoindoline (200 mg, 1.68 mmol) and2-(bromomethyl)oxirane (272 mg, 2.0 mmol) in CH₃CN (10 mL) was addedK₂CO₃ (690 mg, 5 mmol) and reaction mixture was stirred at 25° C. for 16h. The mixture was filtered, concentrated and the crude product was usedin the next step without further purification. (280 mg, yield 95%) MS(ESI⁺) e/z: 176.1 [M+1]⁺.

Step 2: 1-amino-3-(isoindolin-2-yl)propan-2-ol

EtOH (50 mL) was cooled to −78° C. and ammonia gas was bubbled throughthe solution. To the solution was added 2-(oxiran-2-ylmethyl)isoindoline(280 mg, 1.6 mmol), the reaction vessel was sealed and heated at 80° C.for 4 h. The reaction mixture was cooled, concentrated and the crudeproduct was used in the next step without further purification. (300 mg,yield 98%) MS (ESI⁺) e/z: 193.1 [M+1]⁺.

Step 3: ethyl 2-(quinolin-8-yloxy)acetate

To a solution of quinolin-8-ol (3 g, 0.0207 mol) in MeCN (10 mL) wasadded ethyl bromoacetate (4.12 g, 0.025 mol) and K₂CO₃ (5.75 g, 0.0414mol). The mixture was stirred at 80° C. for 12 h, filtered andconcentrated. The residue was purified by column chromatography. (3.9 g,yield 81%) MS (ESI⁺) e/z: 232.1 [M+1]⁺.

Step 4:N-(2-hydroxy-3-(isoindolin-2-yl)propyl)-2-(quinolin-8-yloxy)acetamide

1-amino-3-(isoindolin-2-yl)propan-2-ol (50 mg, 0.26 mmol) and ethyl2-(quinolin-8-yloxy)acetate (60 mg, 0.26 mmol) were dissolved in EtOH (1mL) and heated under microwave conditions at 120° C. for 0.5 h. Thereaction mixture was concentrated and purified by preparative HPLCpurification. (17.2 mg, yield 18%) MS (ESI⁺) e/z: 378.1 [M+1]⁺. ¹H NMR(MeOD, 400 MHz), δ ppm: 8.95-8.87 (m, 1H), 8.45-8.36 (m, 1H), 7.67-7.56(m, 3H), 7.35-7.25 (m, 1H), 7.24-7.12 (m, 4H), 4.83-4.76 (m, 2H),4.09-4.03 (m, 1H), 4.02-3.95 (m, 4H), 3.60-3.54 (m, 1H), 3.46-3.40 (m,1H), 2.92-2.86 (m, 1H), 2.84-2.78 (m, 1H).

Compound 201-(3-((cyclopentylamino)methyl)phenoxy)-3-(isoindolin-2-yl)propan-2-ol

Step 1: 3-(oxiran-2-ylmethoxy)benzaldehyde

To a solution of 3-hydroxybenzaldehyde (2.0 g, 16.38 mmol) in DMF (30mL) at 0° C. was added NaH (983 mg, 24.57 mmol) in portions. After 0.5h, a solution of 2-(bromomethyl)oxirane (2.69 mg, 19.65 mmol) in DMF (5mL) was added and the reaction mixture was warmed to room temperaturefor 5 h. The mixture was concentrated, dissolved in ethyl acetate andwashed with water. The organic portion was dried over sodium sulfate,filtered and concentrated. The crude product was used in the next stepwithout further purification. (2.1 g, yield 72%) MS (ESI⁺) e/z: 179.1[M+1]⁺.

Step 2: N-(3-(oxiran-2-ylmethoxy)benzyl)cyclopentanamine

To a solution of 3-(oxiran-2-ylmethoxy)benzaldehyde (1.0 g, 5.61 mmol)in MeOH (15 mL) was added cyclopentanamine (502 mg, 5.89 mmol) at roomtemperature. After 4 h, sodium borohydride (318 mg, 8.42 mmol) was addedin portions and the mixture was stirred for another 1 h. The reactionmixture was quenched by adding aqueous 1 N HCl until the pH was adjustedto 4-5. The resulting solution was diluted with ethyl acetate, washedwith water, dried over sodium sulfate, filtered and concentrated. Thecrude product was used in the next step without further purification.(1.1 g, yield 79%) MS (ESI⁺) e/z: 248.2 [M+1]⁺.

Step 3: tert-butyl cyclopentyl(3-(oxiran-2-ylmethoxy)benzyl)carbamate

To a solution of N-(3-(oxiran-2-ylmethoxy)benzyl)cyclopentanamine (1.0g, 4.04 mmol) in THF (30 mL) was added Boc₂O (1.32 g, 6.06 mmol) and TEA(614 mg, 6.06 mmol). The reaction mixture was stirred at roomtemperature for 12 h, concentrated, dissolved in ethyl acetate andwashed with water. The organic extracts were dried over sodium sulfate,filtered and concentrated. The crude product was purified by columnchromatography eluting with 10-30% of ethyl acetate in hexane. (1.2 g,yield 86%) MS (ESI⁺) e/z: 348.2 [M+1]⁺.

Step 4: tert-butylcyclopentyl(3-(2-hydroxy-3-(isoindolin-2-yl)propoxy)benzyl)carbamate

To a solution of tert-butylcyclopentyl(3-(oxiran-2-ylmethoxy)benzyl)carbamate (300 mg, 0.86 mmol)in EtOH (5 mL) was added isoindoline (113 mg, 0.95 mmol). The reactionmixture was heated under microwave conditions at 110° C. for 0.8 h. Themixture was concentrated and the crude product was used in the next stepwithout further purification. (220 mg, yield 55%) MS (ESI⁺) e/z: 467.3[M+1]⁺.

Step5:1-(3-((cyclopentylamino)methyl)phenoxy)-3-(isoindolin-2-yl)propan-2-ol

To a solution of tert-butyl cyclopentyl(3-(2-hydroxy-3-(isoindolin-2-yl)propoxy) benzyl)carbamate (200 mg, 0.43 mmol) in ethyl acetate (10 mL)was added HCl/ethyl acetate (5 mL) at 0° C. After 4 h, the reactionmixture was concentrated and purified by preparative HPLC purification.(90 mg, yield 57%) MS (ESI⁺) e/z: 367.3 [M+1]⁺. ¹H NMR (MeOD, 400 MHz),δ ppm: 7.42-7.37 (m, 5H), 7.15-7.05 (m, 3H), 4.72-4.59 (m, 4H),4.48-4.42 (m, 1H), 4.18 (s, 2H), 4.11-4.08 (m, 2H), 3.74-3.30 (m, 3H),2.19-2.12 (m, 2H), 1.84-1.65 (m, 6H).

Biological Assays PRMT5 Biochemical Assay

General Materials. S-adenosylmethionine (SAM), S-adenosylhomocysteine(SAH), bicine, KCl, Tween20, dimethylsulfoxide (DMSO), bovine skingelatin (BSG), and Tris(2-carboxyethyl)phosphine hydrochloride solution(TCEP) were purchased from Sigma-Aldrich at the highest level of puritypossible. ³H-SAM was purchase from American Radiolabeled Chemicals witha specific activity of 80 Ci/mmol. 384-well streptavidin Flashplateswere purchased from PerkinElmer.

Substrates.

Peptide representative of human histone H4 residues 1-15 was synthesizedwith a C-terminal linker-affinity tag motif and a C-terminal amide capby 21^(st) Century Biochemicals. The peptide was high high-perfomanceliquid chromatography (HPLC) purified to greater than 95% purity andconfirmed by liquid chromatography mass spectrometry (LC-MS). Thesequence was Ac-SGRGKGGKGLGKGGA[K-Biot]-amide (SEQ ID NO.:3).

Molecular Biology:

Full-length human PRMT5 (NM_(—)006109.3) transcript variant 1 clone wasamplified from a fetal brain cDNA library, incorporating flanking 5′sequence encoding a FLAG tag (MDYKDDDDK) (SEQ ID NO.:4) fused directlyto Ala 2 of PRMT5. Full-length human MEP50 (NM_(—)024102) clone wasamplified from a human testis cDNA library incorporating a 5′ sequenceencoding a 6-histidine tag (MHHHHHH) (SEQ ID NO.:5) fused directly toArg 2 of MEP50. The amplified genes were sublconed into pENTR/D/TEV(Life Technologies) and subsequently transferred by Gateway™ attL×attRrecombination to pDEST8 baculvirus expression vector (LifeTechnologies).

Protein Expression.

Recombinant baculovirus and Baculovirus-Infected Insect Cells (BIIC)were generated according to Bac-to-Bac kit instructions (LifeTechnologies) and Wasilko, 2006, respectively. Protein over-expressionwas accomplished by infecting exponentially growing Spodopterafrugiperda (SF9) cell culture at 1.2×10⁶ cell/ml with a 5000 folddilution of BIIC stock. Infections were carried out at 27° C. for 72hours, harvested by centrifugation, and stored at −80° C. forpurification.

Protein Purification.

Expressed full-length human Flag-PRMT5/6His-MeP50 protein complex waspurified from cell paste by NiNTA agarose affinity chromatography aftera five hour equilibration of the resin with buffer containing 50 mMTris-HCL, pH 8.0, 25 mM NaCl, and 1 mM TCEP at 4° C., to minimize theadsorption of tubulin impurity by the resin. Flag-PRMT5/6His-MeP50 waseluted with 300 mM Imidazole in the same buffer. The purity of recoveredprotein was 87%. Reference: Wasilko, D. J. and S. E. Lee: “TIPS:titerless infected-cells preservation and scale-up” Bioprocess J., 5(2006), pp. 29-32.

Predicted Translations:

Flag-PRMT5 (SEQ ID NO.: 6)MDYKDDDDKA AMAVGGAGGS RVSSGRDLNC VPEIADTLGA VAKQGFDFLC MPVFHPRFKREFIQEPAKNR PGPQTRSDLL LSGRDWNTLI VGKLSPWIRP DSKVEKIRRN SEAAMLQELNFGAYLGLPAF LLPLNQEDNT NLARVLTNHI HTGHHSSMFW MRVPLVAPED LRDDIIENAPTTHTEEYSGE EKTWMWWHNF RTLCDYSKRI AVALEIGADL PSNHVIDRWL GEPIKAAILPTSIFLTNKKG FPVLSKMHQR LIFRLLKLEV QFIITGTNHH SEKEFCSYLQ YLEYLSQNRPPPNAYELFAK GYEDYLQSPL QPLMDNLESQ TYEVFEKDPI KYSQYQQAIY KCLLDRVPEEEKDTNVQVLM VLGAGRGPLV NASLRAAKQA DRRIKLYAVE KNPNAVVTLE NWQFEEWGSQVTVVSSDMRE WVAPEKADII VSELLGSFAD NELSPECLDG AQHFLKDDGV SIPGEYTSFLAPISSSKLYN EVRACREKDR DPEAQFEMPY VVRLHNFHQL SAPQPCFTFS HPNRDPMIDNNRYCTLEFPV EVNTVLHGFA GYFETVLYQD ITLSIRPETH SPGMFSWFPI LFPIKQPITVREGQTICVRF WRCSNSKKVW YEWAVTAPVC SAIHNPTGRS YTIG L 6His-MEP50(SEQ ID NO.: 7)MHHHHHHRKE TPPPLVPPAA REWNLPPNAP ACMERQLEAA RYRSDGALLL GASSLSGRCWAGSLWLFKDP CAAPNEGFCS AGVQTEAGVA DLTWVGERGI LVASDSGAVE LWELDENETLIVSKFCKYEH DDIVSTVSVL SSGTQAVSGS KDICIKVWDL AQQVVLSSYR AHAAQVTCVAASPHKDSVFL SCSEDNRILL WDTRCPKPAS QIGCSAPGYL PTSLAWHPQQ SEVFVFGDENGTVSLVDTKS TSCVLSSAVH SQCVTGLVFS PHSVPFLASL SEDCSLAVLD SSLSELFRSQAHRDFVRDAT WSPLNHSLLT TVGWDHQVVH HVVPTEPLPA PGPASVTE

General Procedure for PRMT5/MEP50 Enzyme Assays on Peptide Substrates.

The assays were all performed in a buffer consisting of 20 mM Bicine(pH=7.6), 1 mM TCEP, 0.005% BSG, and 0.002% Tween20, prepared on the dayof use. Compounds in 100% DMSO (1 ul) were spotted into a polypropylene384-well V-bottom plates (Greiner) using a Platemate Plus outfitted witha 384-channel head (Thermo Scientific). DMSO (1 ul) was added to Columns11, 12, 23, 24, rows A-H for the maximum signal control and 1 ul of SAH,a known product and inhibitor of PRMT5/MEP50, was added to columns 11,12, 23, 24, rows I-P for the minimum signal control. A cocktail (40 ul)containing the PRMT5/MEP50 enzyme and the peptide was added by MultidropCombi (Thermo-Fisher). The compounds were allowed to incubate withPRMT5/MEP50 for 30 min at 25 degrees Celsius, then a cocktail (10 ul)containing ³H-SAM was added to initiate the reaction (final volume=51ul). The final concentrations of the components were as follows:PRMT5/MEP50 was 4 nM, ³H-SAM was 75 nM, peptide was 40 nM, SAH in theminimum signal control wells was 100 uM, and the DMSO concentration was1%. The assays were stopped by the addition of non-radioactive SAM (10ul) to a final concentration of 600 uM, which dilutes the ³H-SAM to alevel where its incorporation into the peptide substrate is no longerdetectable. 50 ul of the reaction in the 384-well polypropylene platewas then transferred to a 384-well Flashplate and the biotinylatedpeptides were allowed to bind to the streptavidin surface for at least 1hour before being washed three times with 0.1% Tween20 in a BiotekELx405 plate washer. The plates were then read in a PerkinElmer TopCountplate reader to measure the quantity of ³H-labeled peptide bound to theFlashplate surface, measured as disintegrations per minute (dpm) oralternatively, referred to as counts per minute (cpm).

% Inhibition Calculation

${\% \mspace{14mu} {inh}} = {100 - {( \frac{{dpm}_{cmpd} - {dpm}_{\min}}{{dpm}_{\max} - {dpm}_{\min}} ) \times 100}}$

Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.

Four-Parameter IC₅₀ Fit

$Y = {{Bottom} + {m\frac{( {{Top} - {Bottom}} )}{( {1 + ( \frac{X}{{IC}_{50}} )^{{Hill}\mspace{14mu} {Coefficient}}} }}}$

Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.

Z-138 Methylation Assay

Z-138 suspension cells were purchased from ATCC (American Type CultureCollection, Manassas, Va.). RPMI/Glutamax medium,penicillin-streptomycin, heat inactivated fetal bovine serum, and D-PBSwere purchased from Life Technologies, Grand Island, N.Y., USA. Odysseyblocking buffer, 800CW goat anti-rabbit IgG (H+L) antibody, and LicorOdyssey infrared scanner were purchased from Licor Biosciences, Lincoln,Nebr., USA. Symmetric di-methyl arginine antibody was purchased from EMDMillipore, Billerica, Mass., USA. 16% Paraformaldehyde was purchasedfrom Electron Microscopy Sciences, Hatfield, Pa., USA.

Z-138 suspension cells were maintained in growth medium (RPMI 1640supplemented with 10% v/v heat inactivated fetal bovine serum and 100units/mL penicillin-streptomycin) and cultured at 37° C. under 5% CO₂.

Cell Treatment, in Cell Western (ICW) for Detection of SymmetricDi-Methyl Arginine and DNA Content.

Z-138 cells were seeded in assay medium at a concentration of 50,000cells per mL to a 384-well cell culture plate with 50 μL per well.Compound (100 nL) from 384 well source plates was added directly to 384well cell plate. Plates were incubated at 37° C., 5% CO₂ for 96 hours.After four days of incubation, 40 μL of cells from incubated plates wereadded to poly-D-lysine coated 384 well culture plates (BD Biosciences356697). Plates were incubated at room temperature for 30 minutes thenincubated at 37° C., 5% CO₂ for 5 hours. After the incubation, 40 μL perwell of 8% paraformaldehyde in PBS (16% paraformaldahyde was diluted to8% in PBS) was added to each plate and incubated for 30 minutes. Plateswere transferred to a Biotek 405 plate washer and washed 5 times with100 μL per well of wash buffer (1×PBS with 0.1% Triton X-100 (v/v)).Next 30 μL per well of Odyssey blocking buffer were added to each plateand incubated 1 hour at room temperature. Blocking buffer was removedand 20 μL per well of primary antibody was added (symmetric di-methylarginine diluted 1:100 in Odyssey buffer with 0.1% Tween 20 (v/v)) andplates were incubated overnight (16 hours) at 4° C. Plates were washed 5times with 100 μL per well of wash buffer. Next 20 μL per well ofsecondary antibody was added (1:200 800CW goat anti-rabbit IgG (H+L)antibody, 1:1000 DRAQ5 (Biostatus limited) in Odyssey buffer with 0.1%Tween 20 (v/v)) and incubated for 1 hour at room temperature. The plateswere washed 5 times with 100 μL per well wash buffer then 1 time with100 μL per well of water. Plates were allowed to dry at room temperaturethen imaged on the Licor Odyssey machine which measures integratedintensity at 700 nm and 800 nm wavelengths. Both 700 and 800 channelswere scanned.

Calculations:

First, the ratio for each well was determined by:

$( \frac{{symmetric}\mspace{14mu} {di}\text{-}{methyl}\mspace{14mu} {Arginine}\mspace{14mu} 800\mspace{14mu} {nm}\mspace{14mu} {value}}{{DRAQS}\mspace{14mu} 700\mspace{14mu} {nm}\mspace{14mu} {value}} )$

Each plate included fourteen control wells of DMSO only treatment(minimum inhibition) as well as fourteen control wells for maximuminhibition treated with 3 μM of a reference compound (Background wells).The average of the ratio values for each control type was calculated andused to determine the percent inhibition for each test well in theplate. Reference compound was serially diluted three-fold in DMSO for atotal of nine test concentrations, beginning at 3 μM. Percent inhibitionwas determined and IC₅₀ curves were generated using triplicate wells perconcentration of compound.

${{Percent}\mspace{14mu} {Inhibition}} = {100 - ( {( \frac{( {{Individual}\mspace{14mu} {Test}\mspace{14mu} {Sample}\mspace{14mu} {Ratio}} ) - ( {{Background}\mspace{14mu} {Avg}\mspace{14mu} {Ratio}} )}{( {{Minimum}\mspace{14mu} {Inhibition}\mspace{14mu} {Ratio}} ) - ( {{Background}\mspace{14mu} {Average}\mspace{14mu} {Ratio}} )} )*100} )}$

Z-138 Proliferation Assay

Z-138 suspension cells were purchased from ATCC (American Type CultureCollection, Manassas, Va.). RPMI/Glutamax medium,penicillin-streptomycin, heat inactivated fetal bovine serum werepurchased from Life Technologies, Grand Island, N.Y., USA. V-bottompolypropylene 384-well plates were purchased from Greiner Bio-One,Monroe, N.C., USA. Cell culture 384-well white opaque plates werepurchased from Perkin Elmer, Waltham, Mass., USA. Cell-Titer Glo® waspurchased from Promega Corporation, Madison, Wis., USA. SpectraMax M5plate reader was purchased from Molecular Devices LLC, Sunnyvale,Calif., USA.

Z-138 suspension cells were maintained in growth medium (RPMI 1640supplemented with 10% v/v heat inactivated fetal bovine serum andcultured at 37° C. under 5% CO₂. Under assay conditions, cells wereincubated in assay medium (RPMI 1640 supplemented with 10% v/v heatinactivated fetal bovine serum and 100 units/mL penicillin-streptomycin)at 37° C. under 5% CO₂.

For the assessment of the effect of compounds on the proliferation ofthe Z-138 cell line, exponentially growing cells were plated in 384-wellwhite opaque plates at a density of 10,000 cells/ml in a final volume of50 μl of assay medium. A compound source plate was prepared byperforming triplicate nine-point 3-fold serial dilutions in DMSO,beginning at 10 mM (final top concentration of compound in the assay was20 μM and the DMSO was 0.2%). A 100 nL aliquot from the compound stockplate was added to its respective well in the cell plate. The 100%inhibition control consisted of cells treated with 200 nM finalconcentration of staurosporine and the 0% inhibition control consistedof DMSO treated cells. After addition of compounds, assay plates wereincubated for 5 days at 37° C., 5% CO₂, relative humidity>90%. Cellviability was measured by quantitation of ATP present in the cellcultures, adding 35 μl of Cell Titer Glo® reagent to the cell plates.Luminescence was read in the SpectraMax M5 microplate reader. Theconcentration of compound inhibiting cell viability by 50% wasdetermined using a 4-parametric fit of the normalized dose responsecurves.

Results for certain compounds described herein are shown in Table 2.

TABLE 2 Biological Assay Results Cmpd No Biochemical IC₅₀ ICW EC₅₀Proliferation EC₅₀ 1 C — — 2 C — — 3 C — — 4 A B C 5 A B C 6 * — — 7 B B** 8 B B — 9 C — — 10 B B ** 11 B C — 12 B B — 13 B C ** 14 D — — 15 A BC 16 C — — 17 C — — 18 C — — 19 B C ** 20 C — D 21 B B ** 22 B C — 23 AA B 24 A — — 25 D — — 26 B B ** 27 B C ** 28 A B D 29 A B C 30 A B C 31— C — 32 — C ** 33 — B ** 34 — — — For Table 2, “A” indicates an IC₅₀ orEC₅₀ < 0.100 μM, “B” indicates an IC₅₀ or EC₅₀ of 0.101-1.000 μM, “C”indicates an IC₅₀ or EC₅₀ of 1.001-10.000 μM, “D” indicates an IC₅₀ orEC₅₀ of 10.001-50 μM, and “E” indicates an IC₅₀ or EC₅₀ > 50 μM. “—”indicates no data shown. “*” indicates an IC₅₀ or EC₅₀ > 10 μM. “**”indicates an IC₅₀ or EC₅₀ > 20 μM.

OTHER EMBODIMENTS

The foregoing has been a description of certain non-limiting embodimentsof the invention. Those of ordinary skill in the art will appreciatethat various changes and modifications to this description may be madewithout departing from the spirit or scope of the present invention, asdefined in the following claims.

1. A compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein R¹ is hydrogen,R^(z), or —C(O)R^(z), wherein R^(z) is optionally substituted C₁₋₆alkyl; L_(z) is a linker selected from the group consisting of—X_(A)—C(R^(2A))(R^(3A))C(═O)N(R)—, L_(B), or L_(D) wherein: X_(A) is abond, —O—, —N(R)—, —CR^(4A)R^(5A)—, —O—CR^(4A)R^(5A),—N(R)—CR^(4A)R^(5A)—, —O—CR^(4A)R^(5A)—O—, —N(R)—CR^(4A)R^(5A)—O,—N(R)—CR^(4A)R^(5A)—N(R)—, —O—CR^(4A)R^(5A)—N(R)—, —CR^(4A)R^(5A)—O—,—CR^(4A)R^(5A)—N(R)—, —O—CR^(4A)R^(5A)—CR^(6A)R^(7A)—,—N(R)—CR^(4A)R^(5A)—CR^(6A)R^(7A)—, —CR^(6A)R^(7A)—CR^(4A)R^(5A)—O—,—CR^(6A)R^(7A)—CR^(4A)R^(5A)—N(R)—, or —CR^(6A)R^(7A)—CR^(4A)R^(5A)—;each R is independently hydrogen or optionally substituted C₁₋₆aliphatic; R^(2A) and R^(3A) are independently selected from the groupconsisting of hydrogen, halo, —CN, —NO₂, optionally substitutedaliphatic, optionally substituted carbocyclyl, optionally substitutedphenyl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A),—OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B)); or R^(2A) and R^(3A) are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring; R^(4A) and R^(5A) are independentlyselected from the group consisting of hydrogen, halo, —CN, —NO₂,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted phenyl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), —N(R^(B))₇, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R^(4A) and R^(5A) are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring; R^(6A) and R^(7A) are independentlyselected from the group consisting of hydrogen, halo, —CN, —NO₂,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted phenyl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), —N(R^(B)), —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R^(6A) and R^(7A) are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring; each R^(A) is independently selectedfrom the group consisting of hydrogen, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, and optionally substituted heteroaryl; eachR^(B) is independently selected from the group consisting of hydrogen,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted heterocyclyl, optionally substituted aryl, andoptionally substituted heteroaryl, or two R^(B) groups are takentogether with their intervening atoms to form an optionally substitutedheterocyclic ring; L_(B) is —N(R)C(O)—, —C(O)N(R)—, —N(R)C(O)N(R)—,—N(R)C(O)O—, or —OC(O)N(R)—; L_(D) is —O—, —N(R)—, —C(R^(2A))(R^(3A))—,—O—CR^(2A)R^(3A), —N(R)—CR^(2A)R^(3A)—, —O—CR^(2A)R^(3A)—O—,—N(R)—CR^(2A)R^(3A)—O, —N(R)—CR^(2A)R^(3A)—N(R)—,—O—CR^(2A)R^(3A)—N(R)—, —CR^(2A)R^(3A)—O—, —CR^(2A)R^(3A)—N(R)—,—O—CR^(2A)R^(3A)—CR⁹R¹⁰—, —N(R)—CR^(2A)R^(3A)—CR⁹R¹⁰—,—CR^(2A)R^(3A)—CR⁹R¹⁰—, —CR^(2A)R^(3A)—CR⁹R¹⁰—N(R)—, or—CR^(2A)R^(3A)—CR⁹R¹⁰—; R⁹ and R¹⁰ are independently selected from thegroup consisting of hydrogen, halo, —CN, —NO₂, optionally substitutedaliphatic, optionally substituted carbocyclyl; optionally substitutedphenyl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A),—OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁹ and R¹⁰ are taken togetherwith their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring; Ring Z is an optionally substituted,monocyclic or bicyclic, saturated, partially unsaturated, or aromaticring having 0-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur; R²¹, R²², R²³, and R²⁴ are independently hydrogen,halo, or optionally substituted aliphatic; each R^(x) is independentlyselected from the group consisting of halo, —CN, optionally substitutedaliphatic, and —OR′; R′ is hydrogen or optionally substituted aliphatic;and n is 0, 1, 2, 3, 4, 5, 6, 7, or
 8. 2. The compound of claim 1 ofFormula (I^(A)):

or a pharmaceutically acceptable salt thereof, wherein R¹ is hydrogen,R^(z), or —C(O)R^(z), wherein R^(z) is optionally substituted C₁₋₆alkyl; X_(A) is a bond, —O—, —N(R)—, —CR^(4A)R^(5A)—, —O—CR^(4A)R^(5A),—N(R)—CR^(4A)R^(5A)—, —O—CR^(4A)R^(5A)—, —N(R)—CR^(4A)R^(5A)—O,—N(R)—CR^(4A)R^(5A)—N(R)—, —O—CR^(4A)R^(5A)—N(R)—, —CR^(4A)R^(5A)—O—,—CR^(4A)R^(5A)—N(R)—, —O—CR^(4A)R^(5A)—CR^(6A)R^(7A)—,—N(R)—CR^(4A)R^(5A)—CR^(6A)R^(7A)—, —CR^(6A)R^(7A)—CR^(4A)R^(5A)—O—,—CR^(6A)R^(7A)—, —CR^(4A)R^(5A)—O—, —CR^(6A)R^(7A)—CR^(4A)R^(5A)—N(R)—,or —CR^(6A)R^(7A)—CR^(4A)R^(5A)—; each R is independently hydrogen oroptionally substituted C₁₋₆ aliphatic; R^(2A) and R^(3A) areindependently selected from the group consisting of hydrogen, halo, —CN,—NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted phenyl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R^(2A) and R^(3A) are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring; R^(4A) and R^(5A) are independentlyselected from the group consisting of hydrogen, halo, —CN, —NO₂,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted phenyl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R^(4A) and R^(5A) are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring; R^(6A) and R^(7A) are independentlyselected from the group consisting of hydrogen, halo, —CN, —NO₂,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted phenyl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R^(6A) and R^(7A) are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring; each R^(A) is independently selectedfrom the group consisting of hydrogen, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, and optionally substituted heteroaryl; eachR^(B) is independently selected from the group consisting of hydrogen,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted heterocyclyl, optionally substituted aryl, andoptionally substituted heteroaryl, or two R^(B) groups are takentogether with their intervening atoms to form an optionally substitutedheterocyclic ring; R^(8A), R^(9A), R^(10A), and R^(11A) areindependently hydrogen, halo, or optionally substituted aliphatic;Cy^(A) is a monocyclic or bicyclic, saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, wherein Cy^(A) is substituted with 0, 1,2, 3, or 4 R^(y) groups; each R^(y) is independently selected from thegroup consisting of halo, —CN, —NO₂, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted aryl,optionally substituted heterocyclyl, optionally substituted heteroaryl,—OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; each R^(x)is independently selected from the group consisting of halo, —CN,optionally substituted aliphatic, and —OR′; R′ is hydrogen or optionallysubstituted aliphatic; and n is 0, 1, 2, 3, 4, 5, 6, 7, or
 8. 3-93.(canceled)
 94. The compound of claim 1 of Formula (I^(B)):

or a pharmaceutically acceptable salt thereof, wherein R¹ is hydrogen,R^(z), or —C(O)R^(z), wherein R^(z) is optionally substituted C₁₋₆alkyl; L_(B) is —N(R)C(O)—, —C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)O—, or—OC(O)N(R)—; each R is independently hydrogen or optionally substitutedC₁₋₆ aliphatic; Ar is a monocyclic or bicyclic aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein Ar is substituted with 0, 1, 2, 3, 4, or 5 R^(y) groups, asvalency permits; each R^(y) is independently selected from the groupconsisting of halo, —CN, —NO₂, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted aryl,optionally substituted heterocyclyl, optionally substituted heteroaryl,—OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; each R^(A)is independently selected from the group consisting of hydrogen,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted heterocyclyl, optionally substituted aryl, andoptionally substituted heteroaryl; each R^(B) is independently selectedfrom the group consisting of hydrogen, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, and optionally substituted heteroaryl, ortwo R^(B) groups are taken together with their intervening atoms to forman optionally substituted heterocyclic ring; R^(5B), R^(6B), R^(7B), andR^(8B) are independently hydrogen, halo, or optionally substitutedaliphatic; each R^(x) is independently selected from the groupconsisting of halo, —CN, optionally substituted aliphatic, and —OR′; R′is hydrogen or optionally substituted aliphatic; and n is 0, 1, 2, 3, 4,5, 6, 7, or
 8. 95-100. (canceled)
 101. The compound of claim 94, whereinthe compound is of Formula (II^(B)):

or a pharmaceutically acceptable salt thereof. 102-103. (canceled) 104.The compound of claim 94, wherein R¹ is hydrogen.
 105. The compound ofclaim 94, wherein n is 0, 1 or
 2. 106-108. (canceled)
 109. The compoundof claim 94, wherein Ar is heteroaryl. 110-112. (canceled)
 113. Thecompound of claim 94, wherein Ar is substituted with 1 or 2 R^(y)groups. 114-134. (canceled)
 135. The compound of claim 94, wherein atleast one R^(y) is —N(R^(B))₂. 136-149. (canceled)
 150. A compound ofFormula (I^(C)):

or a pharmaceutically acceptable salt thereof, wherein Ring C is anoptionally substituted, 5- to 12-membered, monocyclic or bicyclic,heterocyclyl or heteroaryl having 1-4 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur; R¹ is hydrogen, R^(z), or —C(O)R^(z),wherein R^(z) is optionally substituted C₁₋₆ alkyl; Y is O or S; R^(5B),R^(6B), R^(7B), and R^(8B) are independently hydrogen, halo, oroptionally substituted aliphatic; each R^(x) is independently selectedfrom the group consisting of halo, —CN, optionally substitutedaliphatic, and —OR′; R′ is hydrogen or optionally substituted aliphatic;and n is 0, 1, 2, 3, 4, 5, 6, 7, or
 8. 151-214. (canceled)
 215. Thecompound of claim 1 of formula (I^(D)):

or a pharmaceutically acceptable salt thereof, wherein R¹ is hydrogen,R^(z), or —C(O)R^(z), wherein R^(z) is optionally substituted C₁₋₆alkyl; L_(D) is —O—, —N(R)—, —C(R^(2A))(R^(3A))—, —O—CR^(2A)R^(3A),—N(R)—CR^(2A)R^(3A)—, —O—CR^(2A)R^(3A)—O—, —N(R)—CR^(2A)R^(3A)—O,—N(R)—CR^(2A)R^(3A)—N(R)—, —O—CR^(2A)R^(3A)—N(R)—, —CR^(2A)R^(3A)—O—,—CR^(2A)R^(3A)—N(R)—, —O—CR^(2A)R^(3A)—CR⁹R¹⁰—, —N(R)—CR^(2A)R^(3A)—CR⁹R—, —CR^(2A)R^(3A)—CR⁹R¹⁰—O—, —CR^(2A)R^(3A)—CR⁹R¹⁰—N(R)—, or—CR^(2A)R^(3A)—CR⁹R¹⁰—; each R is independently hydrogen or optionallysubstituted C₁₋₆ aliphatic; R^(2A) and R^(3A) are independently selectedfrom the group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl; optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R^(2A) and R^(3A) are takentogether with their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring; each R^(A) is independently selectedfrom the group consisting of hydrogen, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, and optionally substituted heteroaryl; eachR^(B) is independently selected from the group consisting of hydrogen,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted heterocyclyl, optionally substituted aryl, andoptionally substituted heteroaryl, or two R^(B) groups are takentogether with their intervening atoms to form an optionally substitutedheterocyclic ring; Ring A is a monocyclic or bicyclic, saturated,partially unsaturated, or aromatic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur; R⁴ is-L₁-Cy^(D); L₁ is a bond, —O—, —S—, —N(R)—, —C(O)—, —C(O)N(R)—,—N(R)C(O)N(R)—, —N(R)C(O)—, —N(R)C(O)O—, —OC(O)N(R)—, —SO₂—, —SO₂N(R)—,—N(R)SO₂—, —OC(O)—, —C(O)O—, or an optionally substituted, straight orbranched, C₁₋₆ aliphatic chain wherein one, two, or three methyleneunits of L₁ are optionally and independently replaced by —O—, —S—,—N(R)—, —C(O)—, —C(O)N(R)—, —N(R)C(O)N(R)—, —N(R)C(O)—, —N(R)C(O)O—,—OC(O)N(R)—, —SO₂—, —SO₂N(R)—, —N(R)SO₂—, —OC(O)—, or —C(O)O—; Cy^(D) isan optionally substituted, monocyclic, bicyclic or tricyclic, saturated,partially unsaturated, or aromatic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur; R^(5B),R^(6B), R^(7B), and R^(8B) are independently hydrogen, halo, oroptionally substituted aliphatic; R⁹ and R¹⁰ are independently selectedfrom the group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl; optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁹ and R¹⁰ are taken togetherwith their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring; each R^(y) is independently selectedfrom the group consisting of halo, —CN, —NO₂, optionally substitutedaliphatic, optionally substituted carbocyclyl; optionally substitutedaryl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A),—OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; each R^(x) is independently selectedfrom the group consisting of halo, —CN, optionally substitutedaliphatic, and —OR′; R′ is hydrogen or optionally substituted aliphatic;n is 0, 1, 2, 3, 4, 5, 6, 7, or 8; m is 0, 1, 2, 3, 4, 5, 6, 7, or 8, asvalency permits; and q is 0 or
 1. 216-285. (canceled)
 286. The compoundof claim 1, wherein the compound is selected from the group consistingof:


287. A pharmaceutical composition comprising a compound of claim 1 or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient.
 288. A kit or packaged pharmaceutical comprising acompound of claim 1 or a pharmaceutically acceptable salt thereof, andinstructions for use thereof. 289-293. (canceled)
 294. A method oftreating a PRMT5-mediated disorder, comprising administering to asubject in need thereof a therapeutically effective amount of a compoundof claim 1 or a pharmaceutically acceptable salt thereof. 295.(canceled)
 296. The method of claim 294, wherein the disorder is cancera metabolic disorder, or a blood disorder.
 297. The method of claim 296,wherein the cancer is hematopoietic cancer, lung cancer, prostatecancer, melanoma, or pancreatic cancer.
 298. (canceled)
 299. The methodof claim 296, wherein the metabolic disorder is diabetes or obesity.300-301. (canceled)
 302. The method of claim 296, wherein the blooddisorder is a hemoglobinopathy.
 303. The method of claim 302, whereinthe blood disorder is sickle cell anemia or β-thalessemia. 304.(canceled)